Biofiles Volume 7 Number 2
Neurodegenerative DiseasesAlzheimers Disease
Huntingtons Disease
Parkinsons Disease
Biofilescontents
Introduction 3
Alzheimers Disease Antibodies Proteins Peptides and Assays 4
Huntingtons Disease Antibodies Proteins and Peptides 9
Parkinsons Disease Antibodies Proteins and Peptides 16
Differential protein expression in rat models using the Panoramareg Neurobiology Array 21
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sigmacombiofiles
Highlights from this issue This issue of Biofiles highlights current
research in the areas of Alzheimerrsquos Parkinsonrsquos and Huntingtonrsquos disease
as representative of major efforts to delineate key events in the development
of neurodegenerative diseases Neurodegenerative diseases affect the central nervous system causing
progressive nervous system dysfunction These debilitating and incurable conditions are characterized by loss of neuronal cell function
and are often associated with atrophy of the affected nervous system structures Products featured in this issue include antibodies proteins peptides and assays which represent a broader set of tools offered to
support basic research in neurodegenerative disease
Coming next issue The next issue of Biofiles highlights Epigenetics the study of stable but
potentially reversible alterations in gene expression that occur
without permanent changes in DNA sequence The significance of epigenetic
changes in the development of cancer autism and other diseases is being increasingly recognized Given this developing mechanistic
understanding the field is attracting investigators interested in diverse aspects of chromatin and chromosome biology
Technical contentCarolyn L CrankshawProduct Specialistcarolyncrankshawsialcom
Dr Eliezer KopfManager Manufacturing eliezerkopfsialcom
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 3
Neurodegenerative diseases affect the central nervous system causing progressive nervous system dysfunction These debilitating and incurable conditions are characterized by loss of neuronal cell function and are often associated with atrophy of the affected nervous system structures An important subset of neurodegenerative disease concerns dementias associated with aging Alzheimers disease (AD) is the most common clinically recognized dementia in aging populations and 43 of people 85 or older are thought to suffer from Alzheimers in the United States Parkinsons disease (PD) another common nervous system disorder associated with the elderly affects 1-3 of the population over 60 United Nations population projections estimate a world population of 400 million people 80 years of age or older by the year 2050 Given the financial societal and personal impact of the burden of these diseases determining causes prevention and treatment has become a major focus of basic and clinical research
Study of the etiology of neurodegenerative diseases shows association with genetic factors to be variable within populations for one disease state Even in the case of Huntingtons disease (HD) which is linked to a specific gene how mutant Huntingtins protein effects downstream symptoms of the disorder including dementia is not fully understood The molecular basis of the effects of genetic variation lifestyle and environmental factors including trauma and infection involves multiple signaling pathways Neuropathological hallmarks of dementia include β-amyloid plaques and
neurofibrillary tangles in AD and Lewy body inclusions in PD However while protein aggregation clearly plays a role in neurodegenerative disease there is evidence these are signatures of neuronal damage and additional causative elements remain to be discerned The role of inflammation is an active area of investigation as is the role of nitric oxide signaling The effects of these and other key events on transcriptional regulation and initiation of apoptosis and neurotoxicity continues to be intensively explored
This brochure highlights current understanding in the areas of Alzheimers disease Parkinsons disease and Huntingtons disease as representative of major research efforts to delineate key events in the development of neurodegenerative diseases The associated products represent a broader set of tools offered to support basic research in neurodegenerative disease and in neuroscience
References1 Alzheimers Disease Fact and Figures Alzheimers
Association 2008 httpwwwalzorgdownloadsFacts_Figures_2011pdf
2 Wright WA Geographic and ethnic variation in Parkin-son disease a population-based study of US Medicare beneficiaries Neuroepidemiology 2010 24 143-151
3 Holmes C et al Long-term effects of Aszlig42 immunisation in Alzheimers disease follow-up of a randomised placebo-controlled phase I trial Lancet 2008 372 216-23
4 Griffin WST Inflammation and neurodegenerative disease Am J Clin Nutr 2006 83 472S-474S
5 Breitner JC et al Extended results of the Alzheimers disease anti-inflammatory prevention trial Alzheimers Dement 2011 7 402-11
6 Zhang L et al Role of nitric oxide in Parkinsons disease Pharmacol Ther 2006 109 33-41
IntroductionCarolyn L CrankshawProduct Specialistcarolyncrankshawsialcom
4
Alzheimers Disease Antibodies Proteins Peptides and Assays
Alzheimerprimes Disease
Alzheimers disease (AD) is the most common cause of dementia in the elderly and is characterized by gradual loss of cognitive functions Hallmark pathohistological findings of AD include widespread neuronal degeneration extracellular amyloid plaques and intracellular neurofibrillary tangles (NFT) Biochemical changes affecting multiple pathways contribute to AD pathology Hyperphosphorylation of Tau (MAPT) causes aggregation contributing to the formation of NFT The protein product of DOCK3 stimulates Tau phosphorylation and also interacts with presenilin proteins components of the γ-secretase complex involved in processing of the amyloid β precursor protein (APP) Genetic and biochemical data support the hypothesis that amyloid-β (Aβ) accumulation and aggregation in the brain contribute to the pathogenesis of AD Aβ is derived from sequential proteolytic processing of APP by β-secretases (BACE1 BACE2) and the γ-secretase complex (APH1 NCSTN PSEN1 PSEN2 PSENEN) The longer Aβ42 form has a higher tendency to aggregate and is more toxic than the shorter Aβ40 form A common feature of most Familial Alzheimers Disease (FAD) mutations is an increase in the generation of Aβ peptides particularly Aβ42 Mutations associated with early-onset FAD are found in the APP gene itself or in the presenilin-1 (PSEN1) and presenilin-2 (PSEN2) genes Another gene associated with early-onset FAD TMED10 encodes a protein which regulates γ-secretase activity Ubiquilin
a ubiquitin-like protein interacts with presenilin-2 and is believed to promote presenilin protein accumulation
FAD genetics and mouse models have shed light on early-onset AD pathogenesis but the vast majority of AD cases occur late in life The 4 allele of the apolipoprotein E (APOE) gene (ApoE e4 variant) is a major risk for late-onset AD (LOAD) compared to the APOE2 and APOE3 variants ApoE mediates binding internalization and catabolism of lipoprotein particles via
interaction with members of the low density lipoprotein receptor (LDLR) family The prototype of this family LDLR has a major role maintaining cholesterol homeostasis ApoE receptors include LDLR LDL receptor related proteins (LRP1 LRP1B LRP2) apoE receptor 2 (ApoER2) and the very low density lipoprotein receptor (VLDLR) The basic functions of apoE in normal brain and the role of apoE in neurodegenerative disease remain unknown It is thought the full length and soluble forms of the
Extracellular Space
Cytoplasm
Amyloid-β
Senile plaque Oxidative stress
Lipidperoxidation
Membranedamage
Neuronal death
Neurobrillarytangles
Destabilizedmicrotubules
Impairedaxonal transport
Membranedamage
Hyperphosphorylated
destabilization ofneuronal calcium levels
P
TAU
p35
p25
CDK5
ERK12
CK12
p38 MAPK
PKA
PKCε
MARK
neCa2+
γ-secretase
β-secretase
AKTCDK5
GSK-3β
Calpain
X
γ-secretase
β-secretase
TAU
CalpChemicalDrug or Toxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
OxidizedProtein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
iT
xicant
r c
microRN
a
a
r ct a
h
tor Peptida
Phosph
Kinase
ndenleaep
Phosh
LigadepeNucRece
el
ipio
io
Transme
riTranscriitoRegulat
tTranslatRegulatoRegulat
T
TranscriRegulatTranscr
lated
t
o
Mutated
Transpo
Other
P
O
Alzheimers Disease Amyloid Processing For this and related interactive pathways see sigmacomadsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 5
apoE receptors alter APP processing and Aβ clearance thus contributing to AD pathogenesis Seladin-1 (DHCR24) a crucial enzyme involved in sterol synthesis is downregulated in regions of the brain affected by AD
Another focus in AD research centers around inflammation Patients who have succumbed to Alzheimers show overexpression of interleukin-1 (IL1A) and the soluble astrocyte inflammatory cytokine S100B Further IL1 induces Tau expression and phosphorylation in rat brain and staining brain sections from Alzheimers patients reveals abundant MAPK1 in the same regions as hyperphosphorylated Tau The contribution of these and other events to the pathophysiology and progression of AD continues to be actively investigated
References1 Griffin WST Inflammation and neurodegenerative
disease Am J Clin Nutr 2006 83 472S-474S2 Li Y et al Interleukin-1 mediates pathological effects
of microglia on tau phosphrylation and on synaptophysin
synthesis in cortical neurons through a p38-MAPK pathway J Neurosci 2003 23 1605-11
3 Griffin WST et al Interleukin-1 mediates Alzheimer and Lewy body pathologies J Neuroinflammation 2006 3 5
4 Goldgaber D et al Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimers disease Science 1987 235 877-880
5 Kang J et al The precursor of Alzheimers disease amyloid A4 protein resembles a cell surface receptor Nature 1987 325 733-736
6 Tanzi R et al Amyloid beta protein gene cDNA mRNA distribution and genetic linkage near the Alzheimer locus Science 1987 235 880-884
7 Tanaka S et al Tissue-specific expression of three types of beta-protein precursor mRNA enhancement of protease inhibitor-harboring types in Alzheimers disease brain Biochem Biophys Res Commun 1989 165 1406-1414
8 Haass C and Selkoe DJ Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide Cell 1993 75 1039-1042
9 Vassar R Beta-secretase cleavage of Alzheimers amyloid precursor protein by the transmembrane aspartic protease BACE Science 1999 286 735-741
10 Yan R et al Membrane-anchored aspartyl protease with Alzheimers disease beta-secretase activity Nature 1999 402 533-537
11 Sinha S et al Purification and cloning of amyloid precursor protein beta-secretase from human brain Nature 1999 402 537-540
12 Price DL and Sisodia SS Mutant genes in familial Alzheimers disease and transgenic models Ann Rev Neurosci 1998 21 479-505
13 Tanzi RE et al The Presenilin genes and their role in early-onset familial Alzheimers disease Alzheimers Disease Rev 1996 1 91-98
14 Schellenberg GD et al Genetic linkage evidence for a familial Alzheimers disease locus on chromosome 14 Science 1992 258 668-671
15 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
16 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
17 Yu G et al Nicastrin modulates presenilin-mediated notchglp-1 signal transduction and betaAPP processing Nature 2000 407 48-54
18 Schenk D et al Alzheimers disease A partner for presenilin Nature 2000 407 34-35
19 Sisodia SS Neuroscience An accomplice for gamma-secretase brought into focus Science 2000 289 2296-2297
20 Usdin TB et al Molecular biology of the vesicular ACh transporter Trends Neurosci 1995 18 218-224
21 Varoqui H and Erickson JD The cytoplasmic tail of the vesicular acetylcholine transporter contains a synaptic vesicle targeting signal J Biol Chem 1998 273 9094-9098
Antibodies for Alzheimerprimes ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)
IHC (p)WB
A3981-25ULA3981-200UL
Monoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)IHC (p)
- A5213-2ML
Anti-APH1A goat - APH1A human humanmouse
rat
ELISA (i)WB
- SAB2500076-100UG
Anti-ApoER2 rabbit - LRP8 human human WB A3481-25ULA3481-200UL
Monoclonal Anti-Apolipoprotein E mouse E6D7 APOE human human ELISA (i)IHCIP
WB
A8599-100UL
Anti-BACE 1 N-Terminus (46-62) rabbit - BACE1 human human WB B0681-2ML
Anti-BACE-2 N-terminus (43-60) rabbit - BACE2 human human IF (i)WB
- B7935-200UL
Anti-m-Calpain (Domain III) Large Subunit
rabbit - Capn3 mouseCAPN3 human
Capn3 rat
humanmouse
rat
WB - C0728-1MG
Anti-Glycogen Synthase Kinase-3β (GSK-3β)
rabbit - GSK3B humanGsk3b rat
humanrat
ARRWB
- G7914-2ML
Anti-LRP1 (C-terminal) rabbit - Lrp1 mouseLRP1 human
Lrp1 rat
humanmouse
rat
IF (i)WBWB
L2170-25ULL2170-200UL
Anti-LRP6 (C-terminal region) rabbit - Lrp6 mouseLRP6 human
human IPWB
L2045-25ULL2045-200UL
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
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Biofilescontents
Introduction 3
Alzheimers Disease Antibodies Proteins Peptides and Assays 4
Huntingtons Disease Antibodies Proteins and Peptides 9
Parkinsons Disease Antibodies Proteins and Peptides 16
Differential protein expression in rat models using the Panoramareg Neurobiology Array 21
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sigmacombiofiles
Highlights from this issue This issue of Biofiles highlights current
research in the areas of Alzheimerrsquos Parkinsonrsquos and Huntingtonrsquos disease
as representative of major efforts to delineate key events in the development
of neurodegenerative diseases Neurodegenerative diseases affect the central nervous system causing
progressive nervous system dysfunction These debilitating and incurable conditions are characterized by loss of neuronal cell function
and are often associated with atrophy of the affected nervous system structures Products featured in this issue include antibodies proteins peptides and assays which represent a broader set of tools offered to
support basic research in neurodegenerative disease
Coming next issue The next issue of Biofiles highlights Epigenetics the study of stable but
potentially reversible alterations in gene expression that occur
without permanent changes in DNA sequence The significance of epigenetic
changes in the development of cancer autism and other diseases is being increasingly recognized Given this developing mechanistic
understanding the field is attracting investigators interested in diverse aspects of chromatin and chromosome biology
Technical contentCarolyn L CrankshawProduct Specialistcarolyncrankshawsialcom
Dr Eliezer KopfManager Manufacturing eliezerkopfsialcom
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 3
Neurodegenerative diseases affect the central nervous system causing progressive nervous system dysfunction These debilitating and incurable conditions are characterized by loss of neuronal cell function and are often associated with atrophy of the affected nervous system structures An important subset of neurodegenerative disease concerns dementias associated with aging Alzheimers disease (AD) is the most common clinically recognized dementia in aging populations and 43 of people 85 or older are thought to suffer from Alzheimers in the United States Parkinsons disease (PD) another common nervous system disorder associated with the elderly affects 1-3 of the population over 60 United Nations population projections estimate a world population of 400 million people 80 years of age or older by the year 2050 Given the financial societal and personal impact of the burden of these diseases determining causes prevention and treatment has become a major focus of basic and clinical research
Study of the etiology of neurodegenerative diseases shows association with genetic factors to be variable within populations for one disease state Even in the case of Huntingtons disease (HD) which is linked to a specific gene how mutant Huntingtins protein effects downstream symptoms of the disorder including dementia is not fully understood The molecular basis of the effects of genetic variation lifestyle and environmental factors including trauma and infection involves multiple signaling pathways Neuropathological hallmarks of dementia include β-amyloid plaques and
neurofibrillary tangles in AD and Lewy body inclusions in PD However while protein aggregation clearly plays a role in neurodegenerative disease there is evidence these are signatures of neuronal damage and additional causative elements remain to be discerned The role of inflammation is an active area of investigation as is the role of nitric oxide signaling The effects of these and other key events on transcriptional regulation and initiation of apoptosis and neurotoxicity continues to be intensively explored
This brochure highlights current understanding in the areas of Alzheimers disease Parkinsons disease and Huntingtons disease as representative of major research efforts to delineate key events in the development of neurodegenerative diseases The associated products represent a broader set of tools offered to support basic research in neurodegenerative disease and in neuroscience
References1 Alzheimers Disease Fact and Figures Alzheimers
Association 2008 httpwwwalzorgdownloadsFacts_Figures_2011pdf
2 Wright WA Geographic and ethnic variation in Parkin-son disease a population-based study of US Medicare beneficiaries Neuroepidemiology 2010 24 143-151
3 Holmes C et al Long-term effects of Aszlig42 immunisation in Alzheimers disease follow-up of a randomised placebo-controlled phase I trial Lancet 2008 372 216-23
4 Griffin WST Inflammation and neurodegenerative disease Am J Clin Nutr 2006 83 472S-474S
5 Breitner JC et al Extended results of the Alzheimers disease anti-inflammatory prevention trial Alzheimers Dement 2011 7 402-11
6 Zhang L et al Role of nitric oxide in Parkinsons disease Pharmacol Ther 2006 109 33-41
IntroductionCarolyn L CrankshawProduct Specialistcarolyncrankshawsialcom
4
Alzheimers Disease Antibodies Proteins Peptides and Assays
Alzheimerprimes Disease
Alzheimers disease (AD) is the most common cause of dementia in the elderly and is characterized by gradual loss of cognitive functions Hallmark pathohistological findings of AD include widespread neuronal degeneration extracellular amyloid plaques and intracellular neurofibrillary tangles (NFT) Biochemical changes affecting multiple pathways contribute to AD pathology Hyperphosphorylation of Tau (MAPT) causes aggregation contributing to the formation of NFT The protein product of DOCK3 stimulates Tau phosphorylation and also interacts with presenilin proteins components of the γ-secretase complex involved in processing of the amyloid β precursor protein (APP) Genetic and biochemical data support the hypothesis that amyloid-β (Aβ) accumulation and aggregation in the brain contribute to the pathogenesis of AD Aβ is derived from sequential proteolytic processing of APP by β-secretases (BACE1 BACE2) and the γ-secretase complex (APH1 NCSTN PSEN1 PSEN2 PSENEN) The longer Aβ42 form has a higher tendency to aggregate and is more toxic than the shorter Aβ40 form A common feature of most Familial Alzheimers Disease (FAD) mutations is an increase in the generation of Aβ peptides particularly Aβ42 Mutations associated with early-onset FAD are found in the APP gene itself or in the presenilin-1 (PSEN1) and presenilin-2 (PSEN2) genes Another gene associated with early-onset FAD TMED10 encodes a protein which regulates γ-secretase activity Ubiquilin
a ubiquitin-like protein interacts with presenilin-2 and is believed to promote presenilin protein accumulation
FAD genetics and mouse models have shed light on early-onset AD pathogenesis but the vast majority of AD cases occur late in life The 4 allele of the apolipoprotein E (APOE) gene (ApoE e4 variant) is a major risk for late-onset AD (LOAD) compared to the APOE2 and APOE3 variants ApoE mediates binding internalization and catabolism of lipoprotein particles via
interaction with members of the low density lipoprotein receptor (LDLR) family The prototype of this family LDLR has a major role maintaining cholesterol homeostasis ApoE receptors include LDLR LDL receptor related proteins (LRP1 LRP1B LRP2) apoE receptor 2 (ApoER2) and the very low density lipoprotein receptor (VLDLR) The basic functions of apoE in normal brain and the role of apoE in neurodegenerative disease remain unknown It is thought the full length and soluble forms of the
Extracellular Space
Cytoplasm
Amyloid-β
Senile plaque Oxidative stress
Lipidperoxidation
Membranedamage
Neuronal death
Neurobrillarytangles
Destabilizedmicrotubules
Impairedaxonal transport
Membranedamage
Hyperphosphorylated
destabilization ofneuronal calcium levels
P
TAU
p35
p25
CDK5
ERK12
CK12
p38 MAPK
PKA
PKCε
MARK
neCa2+
γ-secretase
β-secretase
AKTCDK5
GSK-3β
Calpain
X
γ-secretase
β-secretase
TAU
CalpChemicalDrug or Toxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
OxidizedProtein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
iT
xicant
r c
microRN
a
a
r ct a
h
tor Peptida
Phosph
Kinase
ndenleaep
Phosh
LigadepeNucRece
el
ipio
io
Transme
riTranscriitoRegulat
tTranslatRegulatoRegulat
T
TranscriRegulatTranscr
lated
t
o
Mutated
Transpo
Other
P
O
Alzheimers Disease Amyloid Processing For this and related interactive pathways see sigmacomadsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 5
apoE receptors alter APP processing and Aβ clearance thus contributing to AD pathogenesis Seladin-1 (DHCR24) a crucial enzyme involved in sterol synthesis is downregulated in regions of the brain affected by AD
Another focus in AD research centers around inflammation Patients who have succumbed to Alzheimers show overexpression of interleukin-1 (IL1A) and the soluble astrocyte inflammatory cytokine S100B Further IL1 induces Tau expression and phosphorylation in rat brain and staining brain sections from Alzheimers patients reveals abundant MAPK1 in the same regions as hyperphosphorylated Tau The contribution of these and other events to the pathophysiology and progression of AD continues to be actively investigated
References1 Griffin WST Inflammation and neurodegenerative
disease Am J Clin Nutr 2006 83 472S-474S2 Li Y et al Interleukin-1 mediates pathological effects
of microglia on tau phosphrylation and on synaptophysin
synthesis in cortical neurons through a p38-MAPK pathway J Neurosci 2003 23 1605-11
3 Griffin WST et al Interleukin-1 mediates Alzheimer and Lewy body pathologies J Neuroinflammation 2006 3 5
4 Goldgaber D et al Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimers disease Science 1987 235 877-880
5 Kang J et al The precursor of Alzheimers disease amyloid A4 protein resembles a cell surface receptor Nature 1987 325 733-736
6 Tanzi R et al Amyloid beta protein gene cDNA mRNA distribution and genetic linkage near the Alzheimer locus Science 1987 235 880-884
7 Tanaka S et al Tissue-specific expression of three types of beta-protein precursor mRNA enhancement of protease inhibitor-harboring types in Alzheimers disease brain Biochem Biophys Res Commun 1989 165 1406-1414
8 Haass C and Selkoe DJ Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide Cell 1993 75 1039-1042
9 Vassar R Beta-secretase cleavage of Alzheimers amyloid precursor protein by the transmembrane aspartic protease BACE Science 1999 286 735-741
10 Yan R et al Membrane-anchored aspartyl protease with Alzheimers disease beta-secretase activity Nature 1999 402 533-537
11 Sinha S et al Purification and cloning of amyloid precursor protein beta-secretase from human brain Nature 1999 402 537-540
12 Price DL and Sisodia SS Mutant genes in familial Alzheimers disease and transgenic models Ann Rev Neurosci 1998 21 479-505
13 Tanzi RE et al The Presenilin genes and their role in early-onset familial Alzheimers disease Alzheimers Disease Rev 1996 1 91-98
14 Schellenberg GD et al Genetic linkage evidence for a familial Alzheimers disease locus on chromosome 14 Science 1992 258 668-671
15 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
16 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
17 Yu G et al Nicastrin modulates presenilin-mediated notchglp-1 signal transduction and betaAPP processing Nature 2000 407 48-54
18 Schenk D et al Alzheimers disease A partner for presenilin Nature 2000 407 34-35
19 Sisodia SS Neuroscience An accomplice for gamma-secretase brought into focus Science 2000 289 2296-2297
20 Usdin TB et al Molecular biology of the vesicular ACh transporter Trends Neurosci 1995 18 218-224
21 Varoqui H and Erickson JD The cytoplasmic tail of the vesicular acetylcholine transporter contains a synaptic vesicle targeting signal J Biol Chem 1998 273 9094-9098
Antibodies for Alzheimerprimes ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)
IHC (p)WB
A3981-25ULA3981-200UL
Monoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)IHC (p)
- A5213-2ML
Anti-APH1A goat - APH1A human humanmouse
rat
ELISA (i)WB
- SAB2500076-100UG
Anti-ApoER2 rabbit - LRP8 human human WB A3481-25ULA3481-200UL
Monoclonal Anti-Apolipoprotein E mouse E6D7 APOE human human ELISA (i)IHCIP
WB
A8599-100UL
Anti-BACE 1 N-Terminus (46-62) rabbit - BACE1 human human WB B0681-2ML
Anti-BACE-2 N-terminus (43-60) rabbit - BACE2 human human IF (i)WB
- B7935-200UL
Anti-m-Calpain (Domain III) Large Subunit
rabbit - Capn3 mouseCAPN3 human
Capn3 rat
humanmouse
rat
WB - C0728-1MG
Anti-Glycogen Synthase Kinase-3β (GSK-3β)
rabbit - GSK3B humanGsk3b rat
humanrat
ARRWB
- G7914-2ML
Anti-LRP1 (C-terminal) rabbit - Lrp1 mouseLRP1 human
Lrp1 rat
humanmouse
rat
IF (i)WBWB
L2170-25ULL2170-200UL
Anti-LRP6 (C-terminal region) rabbit - Lrp6 mouseLRP6 human
human IPWB
L2045-25ULL2045-200UL
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Neurodegenerative diseases affect the central nervous system causing progressive nervous system dysfunction These debilitating and incurable conditions are characterized by loss of neuronal cell function and are often associated with atrophy of the affected nervous system structures An important subset of neurodegenerative disease concerns dementias associated with aging Alzheimers disease (AD) is the most common clinically recognized dementia in aging populations and 43 of people 85 or older are thought to suffer from Alzheimers in the United States Parkinsons disease (PD) another common nervous system disorder associated with the elderly affects 1-3 of the population over 60 United Nations population projections estimate a world population of 400 million people 80 years of age or older by the year 2050 Given the financial societal and personal impact of the burden of these diseases determining causes prevention and treatment has become a major focus of basic and clinical research
Study of the etiology of neurodegenerative diseases shows association with genetic factors to be variable within populations for one disease state Even in the case of Huntingtons disease (HD) which is linked to a specific gene how mutant Huntingtins protein effects downstream symptoms of the disorder including dementia is not fully understood The molecular basis of the effects of genetic variation lifestyle and environmental factors including trauma and infection involves multiple signaling pathways Neuropathological hallmarks of dementia include β-amyloid plaques and
neurofibrillary tangles in AD and Lewy body inclusions in PD However while protein aggregation clearly plays a role in neurodegenerative disease there is evidence these are signatures of neuronal damage and additional causative elements remain to be discerned The role of inflammation is an active area of investigation as is the role of nitric oxide signaling The effects of these and other key events on transcriptional regulation and initiation of apoptosis and neurotoxicity continues to be intensively explored
This brochure highlights current understanding in the areas of Alzheimers disease Parkinsons disease and Huntingtons disease as representative of major research efforts to delineate key events in the development of neurodegenerative diseases The associated products represent a broader set of tools offered to support basic research in neurodegenerative disease and in neuroscience
References1 Alzheimers Disease Fact and Figures Alzheimers
Association 2008 httpwwwalzorgdownloadsFacts_Figures_2011pdf
2 Wright WA Geographic and ethnic variation in Parkin-son disease a population-based study of US Medicare beneficiaries Neuroepidemiology 2010 24 143-151
3 Holmes C et al Long-term effects of Aszlig42 immunisation in Alzheimers disease follow-up of a randomised placebo-controlled phase I trial Lancet 2008 372 216-23
4 Griffin WST Inflammation and neurodegenerative disease Am J Clin Nutr 2006 83 472S-474S
5 Breitner JC et al Extended results of the Alzheimers disease anti-inflammatory prevention trial Alzheimers Dement 2011 7 402-11
6 Zhang L et al Role of nitric oxide in Parkinsons disease Pharmacol Ther 2006 109 33-41
IntroductionCarolyn L CrankshawProduct Specialistcarolyncrankshawsialcom
4
Alzheimers Disease Antibodies Proteins Peptides and Assays
Alzheimerprimes Disease
Alzheimers disease (AD) is the most common cause of dementia in the elderly and is characterized by gradual loss of cognitive functions Hallmark pathohistological findings of AD include widespread neuronal degeneration extracellular amyloid plaques and intracellular neurofibrillary tangles (NFT) Biochemical changes affecting multiple pathways contribute to AD pathology Hyperphosphorylation of Tau (MAPT) causes aggregation contributing to the formation of NFT The protein product of DOCK3 stimulates Tau phosphorylation and also interacts with presenilin proteins components of the γ-secretase complex involved in processing of the amyloid β precursor protein (APP) Genetic and biochemical data support the hypothesis that amyloid-β (Aβ) accumulation and aggregation in the brain contribute to the pathogenesis of AD Aβ is derived from sequential proteolytic processing of APP by β-secretases (BACE1 BACE2) and the γ-secretase complex (APH1 NCSTN PSEN1 PSEN2 PSENEN) The longer Aβ42 form has a higher tendency to aggregate and is more toxic than the shorter Aβ40 form A common feature of most Familial Alzheimers Disease (FAD) mutations is an increase in the generation of Aβ peptides particularly Aβ42 Mutations associated with early-onset FAD are found in the APP gene itself or in the presenilin-1 (PSEN1) and presenilin-2 (PSEN2) genes Another gene associated with early-onset FAD TMED10 encodes a protein which regulates γ-secretase activity Ubiquilin
a ubiquitin-like protein interacts with presenilin-2 and is believed to promote presenilin protein accumulation
FAD genetics and mouse models have shed light on early-onset AD pathogenesis but the vast majority of AD cases occur late in life The 4 allele of the apolipoprotein E (APOE) gene (ApoE e4 variant) is a major risk for late-onset AD (LOAD) compared to the APOE2 and APOE3 variants ApoE mediates binding internalization and catabolism of lipoprotein particles via
interaction with members of the low density lipoprotein receptor (LDLR) family The prototype of this family LDLR has a major role maintaining cholesterol homeostasis ApoE receptors include LDLR LDL receptor related proteins (LRP1 LRP1B LRP2) apoE receptor 2 (ApoER2) and the very low density lipoprotein receptor (VLDLR) The basic functions of apoE in normal brain and the role of apoE in neurodegenerative disease remain unknown It is thought the full length and soluble forms of the
Extracellular Space
Cytoplasm
Amyloid-β
Senile plaque Oxidative stress
Lipidperoxidation
Membranedamage
Neuronal death
Neurobrillarytangles
Destabilizedmicrotubules
Impairedaxonal transport
Membranedamage
Hyperphosphorylated
destabilization ofneuronal calcium levels
P
TAU
p35
p25
CDK5
ERK12
CK12
p38 MAPK
PKA
PKCε
MARK
neCa2+
γ-secretase
β-secretase
AKTCDK5
GSK-3β
Calpain
X
γ-secretase
β-secretase
TAU
CalpChemicalDrug or Toxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
OxidizedProtein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
iT
xicant
r c
microRN
a
a
r ct a
h
tor Peptida
Phosph
Kinase
ndenleaep
Phosh
LigadepeNucRece
el
ipio
io
Transme
riTranscriitoRegulat
tTranslatRegulatoRegulat
T
TranscriRegulatTranscr
lated
t
o
Mutated
Transpo
Other
P
O
Alzheimers Disease Amyloid Processing For this and related interactive pathways see sigmacomadsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 5
apoE receptors alter APP processing and Aβ clearance thus contributing to AD pathogenesis Seladin-1 (DHCR24) a crucial enzyme involved in sterol synthesis is downregulated in regions of the brain affected by AD
Another focus in AD research centers around inflammation Patients who have succumbed to Alzheimers show overexpression of interleukin-1 (IL1A) and the soluble astrocyte inflammatory cytokine S100B Further IL1 induces Tau expression and phosphorylation in rat brain and staining brain sections from Alzheimers patients reveals abundant MAPK1 in the same regions as hyperphosphorylated Tau The contribution of these and other events to the pathophysiology and progression of AD continues to be actively investigated
References1 Griffin WST Inflammation and neurodegenerative
disease Am J Clin Nutr 2006 83 472S-474S2 Li Y et al Interleukin-1 mediates pathological effects
of microglia on tau phosphrylation and on synaptophysin
synthesis in cortical neurons through a p38-MAPK pathway J Neurosci 2003 23 1605-11
3 Griffin WST et al Interleukin-1 mediates Alzheimer and Lewy body pathologies J Neuroinflammation 2006 3 5
4 Goldgaber D et al Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimers disease Science 1987 235 877-880
5 Kang J et al The precursor of Alzheimers disease amyloid A4 protein resembles a cell surface receptor Nature 1987 325 733-736
6 Tanzi R et al Amyloid beta protein gene cDNA mRNA distribution and genetic linkage near the Alzheimer locus Science 1987 235 880-884
7 Tanaka S et al Tissue-specific expression of three types of beta-protein precursor mRNA enhancement of protease inhibitor-harboring types in Alzheimers disease brain Biochem Biophys Res Commun 1989 165 1406-1414
8 Haass C and Selkoe DJ Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide Cell 1993 75 1039-1042
9 Vassar R Beta-secretase cleavage of Alzheimers amyloid precursor protein by the transmembrane aspartic protease BACE Science 1999 286 735-741
10 Yan R et al Membrane-anchored aspartyl protease with Alzheimers disease beta-secretase activity Nature 1999 402 533-537
11 Sinha S et al Purification and cloning of amyloid precursor protein beta-secretase from human brain Nature 1999 402 537-540
12 Price DL and Sisodia SS Mutant genes in familial Alzheimers disease and transgenic models Ann Rev Neurosci 1998 21 479-505
13 Tanzi RE et al The Presenilin genes and their role in early-onset familial Alzheimers disease Alzheimers Disease Rev 1996 1 91-98
14 Schellenberg GD et al Genetic linkage evidence for a familial Alzheimers disease locus on chromosome 14 Science 1992 258 668-671
15 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
16 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
17 Yu G et al Nicastrin modulates presenilin-mediated notchglp-1 signal transduction and betaAPP processing Nature 2000 407 48-54
18 Schenk D et al Alzheimers disease A partner for presenilin Nature 2000 407 34-35
19 Sisodia SS Neuroscience An accomplice for gamma-secretase brought into focus Science 2000 289 2296-2297
20 Usdin TB et al Molecular biology of the vesicular ACh transporter Trends Neurosci 1995 18 218-224
21 Varoqui H and Erickson JD The cytoplasmic tail of the vesicular acetylcholine transporter contains a synaptic vesicle targeting signal J Biol Chem 1998 273 9094-9098
Antibodies for Alzheimerprimes ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)
IHC (p)WB
A3981-25ULA3981-200UL
Monoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)IHC (p)
- A5213-2ML
Anti-APH1A goat - APH1A human humanmouse
rat
ELISA (i)WB
- SAB2500076-100UG
Anti-ApoER2 rabbit - LRP8 human human WB A3481-25ULA3481-200UL
Monoclonal Anti-Apolipoprotein E mouse E6D7 APOE human human ELISA (i)IHCIP
WB
A8599-100UL
Anti-BACE 1 N-Terminus (46-62) rabbit - BACE1 human human WB B0681-2ML
Anti-BACE-2 N-terminus (43-60) rabbit - BACE2 human human IF (i)WB
- B7935-200UL
Anti-m-Calpain (Domain III) Large Subunit
rabbit - Capn3 mouseCAPN3 human
Capn3 rat
humanmouse
rat
WB - C0728-1MG
Anti-Glycogen Synthase Kinase-3β (GSK-3β)
rabbit - GSK3B humanGsk3b rat
humanrat
ARRWB
- G7914-2ML
Anti-LRP1 (C-terminal) rabbit - Lrp1 mouseLRP1 human
Lrp1 rat
humanmouse
rat
IF (i)WBWB
L2170-25ULL2170-200UL
Anti-LRP6 (C-terminal region) rabbit - Lrp6 mouseLRP6 human
human IPWB
L2045-25ULL2045-200UL
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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4
Alzheimers Disease Antibodies Proteins Peptides and Assays
Alzheimerprimes Disease
Alzheimers disease (AD) is the most common cause of dementia in the elderly and is characterized by gradual loss of cognitive functions Hallmark pathohistological findings of AD include widespread neuronal degeneration extracellular amyloid plaques and intracellular neurofibrillary tangles (NFT) Biochemical changes affecting multiple pathways contribute to AD pathology Hyperphosphorylation of Tau (MAPT) causes aggregation contributing to the formation of NFT The protein product of DOCK3 stimulates Tau phosphorylation and also interacts with presenilin proteins components of the γ-secretase complex involved in processing of the amyloid β precursor protein (APP) Genetic and biochemical data support the hypothesis that amyloid-β (Aβ) accumulation and aggregation in the brain contribute to the pathogenesis of AD Aβ is derived from sequential proteolytic processing of APP by β-secretases (BACE1 BACE2) and the γ-secretase complex (APH1 NCSTN PSEN1 PSEN2 PSENEN) The longer Aβ42 form has a higher tendency to aggregate and is more toxic than the shorter Aβ40 form A common feature of most Familial Alzheimers Disease (FAD) mutations is an increase in the generation of Aβ peptides particularly Aβ42 Mutations associated with early-onset FAD are found in the APP gene itself or in the presenilin-1 (PSEN1) and presenilin-2 (PSEN2) genes Another gene associated with early-onset FAD TMED10 encodes a protein which regulates γ-secretase activity Ubiquilin
a ubiquitin-like protein interacts with presenilin-2 and is believed to promote presenilin protein accumulation
FAD genetics and mouse models have shed light on early-onset AD pathogenesis but the vast majority of AD cases occur late in life The 4 allele of the apolipoprotein E (APOE) gene (ApoE e4 variant) is a major risk for late-onset AD (LOAD) compared to the APOE2 and APOE3 variants ApoE mediates binding internalization and catabolism of lipoprotein particles via
interaction with members of the low density lipoprotein receptor (LDLR) family The prototype of this family LDLR has a major role maintaining cholesterol homeostasis ApoE receptors include LDLR LDL receptor related proteins (LRP1 LRP1B LRP2) apoE receptor 2 (ApoER2) and the very low density lipoprotein receptor (VLDLR) The basic functions of apoE in normal brain and the role of apoE in neurodegenerative disease remain unknown It is thought the full length and soluble forms of the
Extracellular Space
Cytoplasm
Amyloid-β
Senile plaque Oxidative stress
Lipidperoxidation
Membranedamage
Neuronal death
Neurobrillarytangles
Destabilizedmicrotubules
Impairedaxonal transport
Membranedamage
Hyperphosphorylated
destabilization ofneuronal calcium levels
P
TAU
p35
p25
CDK5
ERK12
CK12
p38 MAPK
PKA
PKCε
MARK
neCa2+
γ-secretase
β-secretase
AKTCDK5
GSK-3β
Calpain
X
γ-secretase
β-secretase
TAU
CalpChemicalDrug or Toxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
OxidizedProtein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
iT
xicant
r c
microRN
a
a
r ct a
h
tor Peptida
Phosph
Kinase
ndenleaep
Phosh
LigadepeNucRece
el
ipio
io
Transme
riTranscriitoRegulat
tTranslatRegulatoRegulat
T
TranscriRegulatTranscr
lated
t
o
Mutated
Transpo
Other
P
O
Alzheimers Disease Amyloid Processing For this and related interactive pathways see sigmacomadsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 5
apoE receptors alter APP processing and Aβ clearance thus contributing to AD pathogenesis Seladin-1 (DHCR24) a crucial enzyme involved in sterol synthesis is downregulated in regions of the brain affected by AD
Another focus in AD research centers around inflammation Patients who have succumbed to Alzheimers show overexpression of interleukin-1 (IL1A) and the soluble astrocyte inflammatory cytokine S100B Further IL1 induces Tau expression and phosphorylation in rat brain and staining brain sections from Alzheimers patients reveals abundant MAPK1 in the same regions as hyperphosphorylated Tau The contribution of these and other events to the pathophysiology and progression of AD continues to be actively investigated
References1 Griffin WST Inflammation and neurodegenerative
disease Am J Clin Nutr 2006 83 472S-474S2 Li Y et al Interleukin-1 mediates pathological effects
of microglia on tau phosphrylation and on synaptophysin
synthesis in cortical neurons through a p38-MAPK pathway J Neurosci 2003 23 1605-11
3 Griffin WST et al Interleukin-1 mediates Alzheimer and Lewy body pathologies J Neuroinflammation 2006 3 5
4 Goldgaber D et al Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimers disease Science 1987 235 877-880
5 Kang J et al The precursor of Alzheimers disease amyloid A4 protein resembles a cell surface receptor Nature 1987 325 733-736
6 Tanzi R et al Amyloid beta protein gene cDNA mRNA distribution and genetic linkage near the Alzheimer locus Science 1987 235 880-884
7 Tanaka S et al Tissue-specific expression of three types of beta-protein precursor mRNA enhancement of protease inhibitor-harboring types in Alzheimers disease brain Biochem Biophys Res Commun 1989 165 1406-1414
8 Haass C and Selkoe DJ Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide Cell 1993 75 1039-1042
9 Vassar R Beta-secretase cleavage of Alzheimers amyloid precursor protein by the transmembrane aspartic protease BACE Science 1999 286 735-741
10 Yan R et al Membrane-anchored aspartyl protease with Alzheimers disease beta-secretase activity Nature 1999 402 533-537
11 Sinha S et al Purification and cloning of amyloid precursor protein beta-secretase from human brain Nature 1999 402 537-540
12 Price DL and Sisodia SS Mutant genes in familial Alzheimers disease and transgenic models Ann Rev Neurosci 1998 21 479-505
13 Tanzi RE et al The Presenilin genes and their role in early-onset familial Alzheimers disease Alzheimers Disease Rev 1996 1 91-98
14 Schellenberg GD et al Genetic linkage evidence for a familial Alzheimers disease locus on chromosome 14 Science 1992 258 668-671
15 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
16 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
17 Yu G et al Nicastrin modulates presenilin-mediated notchglp-1 signal transduction and betaAPP processing Nature 2000 407 48-54
18 Schenk D et al Alzheimers disease A partner for presenilin Nature 2000 407 34-35
19 Sisodia SS Neuroscience An accomplice for gamma-secretase brought into focus Science 2000 289 2296-2297
20 Usdin TB et al Molecular biology of the vesicular ACh transporter Trends Neurosci 1995 18 218-224
21 Varoqui H and Erickson JD The cytoplasmic tail of the vesicular acetylcholine transporter contains a synaptic vesicle targeting signal J Biol Chem 1998 273 9094-9098
Antibodies for Alzheimerprimes ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)
IHC (p)WB
A3981-25ULA3981-200UL
Monoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)IHC (p)
- A5213-2ML
Anti-APH1A goat - APH1A human humanmouse
rat
ELISA (i)WB
- SAB2500076-100UG
Anti-ApoER2 rabbit - LRP8 human human WB A3481-25ULA3481-200UL
Monoclonal Anti-Apolipoprotein E mouse E6D7 APOE human human ELISA (i)IHCIP
WB
A8599-100UL
Anti-BACE 1 N-Terminus (46-62) rabbit - BACE1 human human WB B0681-2ML
Anti-BACE-2 N-terminus (43-60) rabbit - BACE2 human human IF (i)WB
- B7935-200UL
Anti-m-Calpain (Domain III) Large Subunit
rabbit - Capn3 mouseCAPN3 human
Capn3 rat
humanmouse
rat
WB - C0728-1MG
Anti-Glycogen Synthase Kinase-3β (GSK-3β)
rabbit - GSK3B humanGsk3b rat
humanrat
ARRWB
- G7914-2ML
Anti-LRP1 (C-terminal) rabbit - Lrp1 mouseLRP1 human
Lrp1 rat
humanmouse
rat
IF (i)WBWB
L2170-25ULL2170-200UL
Anti-LRP6 (C-terminal region) rabbit - Lrp6 mouseLRP6 human
human IPWB
L2045-25ULL2045-200UL
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Internet sigma-aldrichcom
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 5
apoE receptors alter APP processing and Aβ clearance thus contributing to AD pathogenesis Seladin-1 (DHCR24) a crucial enzyme involved in sterol synthesis is downregulated in regions of the brain affected by AD
Another focus in AD research centers around inflammation Patients who have succumbed to Alzheimers show overexpression of interleukin-1 (IL1A) and the soluble astrocyte inflammatory cytokine S100B Further IL1 induces Tau expression and phosphorylation in rat brain and staining brain sections from Alzheimers patients reveals abundant MAPK1 in the same regions as hyperphosphorylated Tau The contribution of these and other events to the pathophysiology and progression of AD continues to be actively investigated
References1 Griffin WST Inflammation and neurodegenerative
disease Am J Clin Nutr 2006 83 472S-474S2 Li Y et al Interleukin-1 mediates pathological effects
of microglia on tau phosphrylation and on synaptophysin
synthesis in cortical neurons through a p38-MAPK pathway J Neurosci 2003 23 1605-11
3 Griffin WST et al Interleukin-1 mediates Alzheimer and Lewy body pathologies J Neuroinflammation 2006 3 5
4 Goldgaber D et al Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimers disease Science 1987 235 877-880
5 Kang J et al The precursor of Alzheimers disease amyloid A4 protein resembles a cell surface receptor Nature 1987 325 733-736
6 Tanzi R et al Amyloid beta protein gene cDNA mRNA distribution and genetic linkage near the Alzheimer locus Science 1987 235 880-884
7 Tanaka S et al Tissue-specific expression of three types of beta-protein precursor mRNA enhancement of protease inhibitor-harboring types in Alzheimers disease brain Biochem Biophys Res Commun 1989 165 1406-1414
8 Haass C and Selkoe DJ Cellular processing of beta-amyloid precursor protein and the genesis of amyloid beta-peptide Cell 1993 75 1039-1042
9 Vassar R Beta-secretase cleavage of Alzheimers amyloid precursor protein by the transmembrane aspartic protease BACE Science 1999 286 735-741
10 Yan R et al Membrane-anchored aspartyl protease with Alzheimers disease beta-secretase activity Nature 1999 402 533-537
11 Sinha S et al Purification and cloning of amyloid precursor protein beta-secretase from human brain Nature 1999 402 537-540
12 Price DL and Sisodia SS Mutant genes in familial Alzheimers disease and transgenic models Ann Rev Neurosci 1998 21 479-505
13 Tanzi RE et al The Presenilin genes and their role in early-onset familial Alzheimers disease Alzheimers Disease Rev 1996 1 91-98
14 Schellenberg GD et al Genetic linkage evidence for a familial Alzheimers disease locus on chromosome 14 Science 1992 258 668-671
15 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
16 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
17 Yu G et al Nicastrin modulates presenilin-mediated notchglp-1 signal transduction and betaAPP processing Nature 2000 407 48-54
18 Schenk D et al Alzheimers disease A partner for presenilin Nature 2000 407 34-35
19 Sisodia SS Neuroscience An accomplice for gamma-secretase brought into focus Science 2000 289 2296-2297
20 Usdin TB et al Molecular biology of the vesicular ACh transporter Trends Neurosci 1995 18 218-224
21 Varoqui H and Erickson JD The cytoplasmic tail of the vesicular acetylcholine transporter contains a synaptic vesicle targeting signal J Biol Chem 1998 273 9094-9098
Antibodies for Alzheimerprimes ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)
IHC (p)WB
A3981-25ULA3981-200UL
Monoclonal Anti-β-Amyloid mouse BAM-10 APP human human ELISA (i)IHC (p)
- A5213-2ML
Anti-APH1A goat - APH1A human humanmouse
rat
ELISA (i)WB
- SAB2500076-100UG
Anti-ApoER2 rabbit - LRP8 human human WB A3481-25ULA3481-200UL
Monoclonal Anti-Apolipoprotein E mouse E6D7 APOE human human ELISA (i)IHCIP
WB
A8599-100UL
Anti-BACE 1 N-Terminus (46-62) rabbit - BACE1 human human WB B0681-2ML
Anti-BACE-2 N-terminus (43-60) rabbit - BACE2 human human IF (i)WB
- B7935-200UL
Anti-m-Calpain (Domain III) Large Subunit
rabbit - Capn3 mouseCAPN3 human
Capn3 rat
humanmouse
rat
WB - C0728-1MG
Anti-Glycogen Synthase Kinase-3β (GSK-3β)
rabbit - GSK3B humanGsk3b rat
humanrat
ARRWB
- G7914-2ML
Anti-LRP1 (C-terminal) rabbit - Lrp1 mouseLRP1 human
Lrp1 rat
humanmouse
rat
IF (i)WBWB
L2170-25ULL2170-200UL
Anti-LRP6 (C-terminal region) rabbit - Lrp6 mouseLRP6 human
human IPWB
L2045-25ULL2045-200UL
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
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6
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk3 ratMAPK1 humanMapk3 mouseMapk1 mouse
Mapk1 ratMAPK3 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ELISA (i)ICC
IHC (p)IP
WB
- M8159-2ML
Monoclonal Anti-MAP Kinase Activated (Diphosphorylated ERK-1amp2)
mouse MAPK-YT Mapk1 mouseMAPK3 human
Mapk3 ratMapk3 mouse
Mapk1 ratMAPK1 human
Caenorhabditis elegansDrosophila
Xenopusbovine
hamsterhumanmouse
ratyeast
ARRELISA (i)
ICCIHC (p)
IPWB
M9692-200UL
Anti-Nicastrin rabbit - NCSTN human humanmouse
rat
ARRIF (i)
IPWB
N1660-2ML
Anti-p35 (Cdk5 Regulator) rabbit - CDK5R1 humanCdk5r1 rat
humanrat
ARRWB
- P9489-2ML
Anti-Pen-2 rabbit - Psen2 mousePSENEN human
human ARRWB
P5622-200UL
Anti-phospho-PKB (pSer473) rabbit - AKT1 humanAkt1 rat
Akt1 mouse
mouserat
ARRWB
- P4112-2ML
Anti-Presenilin-1 (S182) rabbit - Psen1 mousePsen1 rat
PSEN1 human
Xenopushumanmouse
rat
ARRIHC (p)
WB
P7854-2ML
Anti-Seladin-1 rabbit - DHCR24 human human WB S8571-200UL
Anti-τ (Tau) rabbit - MAPT human chickenwide range
WB T6402-2MLT6402-1ML
Monoclonal Anti-τ (Tau) mouse TAU-2 MAPT human bovinechickenhumanmonkey
ARRIHC (p)
WB
T5530-2MLT5530-5ML
Anti-TMP21 (C-terminal) rabbit - TMED10 humanTmed10 rat
Tmed10 mouse
humanmouse
rat
WB T3827-25ULT3827-200UL
Anti-Ubiquilin-1 rabbit - UBQLN1 human human WB U7258-25ULU7258-200UL
Monoclonal Anti-Vimentin mouse LN-6 Vim ratVim mouseVIM human
bovinefeline
humanmouse
pigrabbit
ratsheep
IF (i)IHC (p)
IPWB
- V2258-2MLV2258-5ML
Checkmark denotes antibodies represented on the Panoramareg Neurobiology Microarray
Antibodies for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 7
Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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Proteins amp Peptides for Alzheimerprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
AKT2 active GST tagged human
AKT2 is a serinethreonine kinase that functions in cellular signaling pathways regulating glucose metabolism transcription survival cell proliferation angiogenesis and cell motility
AKT2 ge70 SDS-PAGE A2233-10UG
Amyloid Precursor Protein α Secreted human
αminussecretase-cleaved soluble amyloid precursor protein has been shown to have neuroprotective properties Several G protein-coupled receptors are known to activate α-secretase-dependent processing of APP
APP gt90 SDS-PAGE S9564-25UG
Amyloid Precursor Protein β Secreted human
Proteolytic cleavage product of amyloid β precursor protein (APP) sAPPβ is thought to modulate neuronal function and cell survival
APP gt85 SDS-PAGE S4316-25UG
Amyloid β Protein Fragment 1-40
β-Amyloid fragment that is neurotoxic in vivo and in vitro in neuronal cell cultures
APP ge90 HPLC A1075-1MGA1075-5MG
Amyloid β Protein Fragment 1-42
The predominant fragment of amyloid β-protein in Alzheimers disease APP ge95 HPLC A9810-1MG
Amyloid β Protein Fragment 25-35
Functional domain of Aβ required for both neurotrophic and neurotoxic effects
APP ge97 HPLC A4559-250UGA4559-1MG
Amyloid β Protein Fragment 1-40 All D-Amino Acids
This D-amino acid peptide functions as a control useful in elucidating structural dependence of aggregation properties characteristic of the amyloid β 1-40 peptide associated with plaque formation and Alzheimers disease
APP gt70 HPLC A5973-5MG
Apolipoprotein E4 human The ApoE4 isoform of ApoE correlates with increased incidence of Alzheimers disease and has been shown to regulate lipid metabolism and bind amyloid β Recombinant ApoE4 retains full biological activity and can be used to study interactions of ApoE4 with amyloid-β Tau and LDLR
APOE ge90 SDS-PAGE and HPLC
A3234-100UG
CDK5p25 active GST tagged human
CDK5 abundant in the mammalian brain is activated upon binding to neuronal protein p35 CDK5p35 breakdown to CDK5p25 is associated with increased neurotoxicity as well as neurodegenerative diseases including Alzheimers and Parkinsons
CDK5CDK5R1
ge70 SDS-PAGE C0745-10UG
ERK1 active untagged human
ERK1 (MAPK3) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK3 ge70 SDS-PAGE E7407-10UG
ERK2 active GST tagged human
ERK2 (MAPK1) participates in cellular signaling cascades that are activated in response to numerous growth factors and cytokines Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
MAPK1 ge70 SDS-PAGE E1283-10UG
Imna(--) mouse embryonic fibroblasts stained with Monoclonal Anti-Vimentin clone LN-6 (Cat No V2258)
From Shyam Khatau Department of Chemical and Biomolecular Engineering Johns Hopkins University Baltimore MD
Drosophila wing imaginal disc (500 micrometers long) was stained with Monoclonal Anti-MAP Kinase Activated (Cat No M8159)
From L Gabay R Seger B-Z Shilo Weizmann Institute of Science ehovot Israel reproduced cover photograph from Science 277 1103 (1997) Used with permission
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
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Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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Internet sigma-aldrichcom
8
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No GSK3β active
His tagged humanGSK3B a serinethreonine kinase functions in physiological processes including the control of glycogen metabolism cell division proliferation motility and survival Current evidence indicates GSK3B plays a role in neurological disease and it is known to phosphorylate both Tau and presenilin-1
GSK3B ge70 SDS-PAGE G4296-10UG
Presenilin-1 N-Terminal Peptide
Used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN1 ge50 HPLC P2490-1MG
Presenilin-2 N-Terminal Peptide
Product used to study production of Aβ[X-42] peptide and accumulation of endogenous presenilin
PSEN2 ge85 HPLC P2740-1MG
Protein Kinase A Catalytic Subunit β Active human
A catalytic subunit of cAMP-dependent protein kinase the protein encoded by PRKACB catalyzes events downstream of GPCRs including cell cycle differentiation and proliferation When activated this subunit acts on metabolic enzymes ion channels and transcription factors such as CREB
PRKACB ge85 SDS-PAGE P6998-5UG
β-Secretase human Transmembrane protease responsible for the β site cleavage of the amyloid precursor protein (APP) to produce amyloid β peptide
BACE1 ge90 SDS-PAGE S4195-50UG
Tau-352 human Isoform of Tau variant 0N3R having 3 microtubule binding repeats (R) and no amino terminal inserts (N)
MAPT ge90 SDS-PAGE T9950-50UG
Tau-412 human Isoform of Tau variant 1N4R having 4 microtubule binding repeats (R) and one amino terminal insert (N)
MAPT ge90 SDS-PAGE T0326-50UG
Tau-441 human Isoform of Tau variant 2N4R having 4 microtubule binding repeats (R) and 2 amino terminal inserts (N)
MAPT ge90 SDS-PAGE T0576-50UG
8 Vimentin His tagged human
Vimentin is a member of the intermediate filament family of proteins that plays a significant role in supporting and anchoring organelles in the cytosol It functions to maintain cell shape and stabilize cytoskeletal interactions
VIM ge90 SDS-PAGE SRP5150-50UG
Assays for Alzheimerprimes ResearchBACE-1 Activity Assay
Product Name Application Cat NoSensiZyme BACE1 Activity Assay Kit sufficient for 96 multiwell tests
The BACE1 Activity Assay Kit provides all the reagents required for highly sensitive detection of BACE1 activity in cell extracts cell culture media tissue extracts and purified enzyme preparations and also for inhibitor screening This assay is both sensitive and specific The enhanced sensitivity is achieved by the signal amplification via the chain reaction The specificity is achieved by both the immunochemical isolation of the BACE1 enzyme from the extract by specific antibodies bound to the 96-well plate and the use of an enzyme substrate (Substrate A) containing a BACE1 specific cleavage site
CS1060-1KT
β-Secretase (BACE1) Activity Detection Kit (Fluorescent) 1 kit sufficient for 250 reactions
The kit provides all the reagents required for an efficient detection of BACE1 activity It contains an enzyme to be used for screening of potential BACE1 inhibitors The assay is based on the fluorescence resonance energy transfer (FRET) method in which the fluorescence signal enhancement is observed after substrate cleavage by BACE1
CS0010-1KT
To view additional products for Alzheimers Disease Research visit sigmacomalz
Proteins amp Peptides for Alzheimerprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 9
Huntingtons disease (HD) is an autosomal dominant late-onset neurodegenerative disorder characterized by a selective neuronal cell death in the cortex and striatum leading to cognitive dysfunction motor impairment and behavioral changes The underlying cause of HD is the expansion of a CAG repeat located within the first exon of the Huntingtin gene (HTT) In persons with HD the HTT gene is found to contain 36 or more CAG repeats resulting in a mutant form of the Huntingtin protein The current hypothesis in HD is that neuronal degeneration results from the combined effects of a gain-of-function in the mutated form of HTT along with a loss of function in the wild-type HTT Pathogenesis in HD appears to involve different mechanisms
1 HD mutation is translated into an expanded polyglutamine tract (polyQ) that induces conformational changes and abnormal folding in the mutated Huntingtin These insoluble proteins accumulate as ubiquitinated cytoplasmic perinuclear aggregates The resulting perinuclear inclusions impair the ubiquitin-proteasome system leading to the accumulation of more misfolded proteins and cell death
2 HTT mutation results in abnormal protein interactions For example mutant Huntingtin interferes with the binding of disks large associated protein 4 (DLGAP4) to the glutamate receptor NMDAR1 (GRIN1) This results in receptor hypersensitivity an influx of Ca2+ and excitotoxicity Additionally increased Ca2+ levels activate caspases leading to cell apoptosis cleavage of mutant Huntingtin and the generation of toxic N-terminal fragments In HD mutant Huntingtin can also inhibit transcription by failing to bind
to the repressor REST in the cytoplasm This results in an accumulation of the repressor in the nucleus and inhibition of brain-derived neurotrophic factor (BDNF) transcription which is an important survival factor for striatal neurons Finally decreased binding between mutant Huntingtin and proteins such as MLK2 (MAP3K10) HIP1 and HIP14 leads to apoptotic cell death impaired vesicle trafficking and endocytosis
3 Huntingtin mutation leads to aggregate sequestration of various proteins including transcription factors Proteolytically cleaved N-terminal fragments of mutated Huntingtin can translocate into the nucleus to form neuronal intranuclear inclusions Once there mutated Huntingtin recruits transcription factors such as CBP (CREBBP EP300) TBP and SIN3A which disrupt gene transcription leading to neurodegeneration
References1 Hu Y et al Bcl-XL interacts with Apaf-1 and inhibits
Apaf-1-dependent caspase-9 activation Proc Natl Acad Sci USA 1998 95 4386-4391
2 Rangone H et al The serum- and glucocorticoid- induced kinase SGK inhibits mutant Huntingtin-induced toxicity by phosphorylating serine 421 of Huntingtin Eur J Neurosci 2004 19 273-279
3 Nakagawa T and Yuan J Cross-talk between two cysteine protease families Activation of caspase-12 by calpain in apoptosis J Cell Biol 2000 150 887-894
4 Heumann R et al Transgenic activation of Ras in neurons promotes hypertrophy and protects from lesion-induced degeneration J Cell Biol 2000 151 1537-1548
5 Weber MM et al Rat somatotroph insulin-like growth factor-II (IGF-II) signaling role of the IGF-I receptor Endocrinology 1992 131 2147-2153
6 Liu YF et al SH3 domain-dependent association of Huntingtin with epidermal growth factor receptor signaling complexes J Biol Chem 1997 272 8121-8124
7 Perkins CL et al The role of Apaf-1 caspase-9 and bid proteins in etoposide- or paclitaxel-induced mitochondrial events during apoptosis Cancer Res 2000 60 1645-1653
8 Tartare-Deckert S et al Interaction of the molecular
weight 85K regulatory subunit of the phosphatidylino-sitol 3-kinase with the insulin receptor and the insulin-like growth factor-1 (IGF- I) receptor comparative study using the yeast two-hybrid system Endocrinology 1996 137 1019-1024
9 Doonan F et al Caspase-Independent Photoreceptor Apoptosis in Mouse Models of Retinal Degeneration J Neurosci 2003 23 5723-5731
10 Liu YF et al Activation of MLK2-mediated signaling cascades by polyglutamine-expanded Huntingtin J Biol Chem 2000 275 19035-19040
11 Borg JP et al The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein Mol Cell Biol 1996 16 6229-6241
12 Petrosillo G et al Ca2+-induced Reactive Oxygen Species Production Promotes Cytochrome c Release from Rat Liver Mitochondria via Mitochondrial Permeability Transition (MPT)-dependent and MPT-independent Mechanisms role of cardiolipin J Biol Chem 2004 279 53103-53108
13 Adler V et al Complexes of p21RAS with JUN N-terminal kinase and JUN proteins Proc Natl Acad Sci USA 1995 92 10585-10589
14 Thien CB and Langdon WY Tyrosine kinase activity of the EGF receptor is enhanced by the expression of oncogenic 70Z-Cbl Oncogene 1997 15 2909-2919
15 Yazgan O and Pfarr CM Regulation of two JunD isoforms by Jun-N-terminal kinases J Biol Chem 2002 277 29710-29718
16 Hirai S et al MSTMLK2 a member of the mixed lineage kinase family directly phosphorylates and activates SEK1 an activator of c-Jun N-terminal kinasestress-activated protein kinase J Biol Chem 1997 272 15167-15173
17 Hattori S et al Activation of mitogen-activated protein kinase and its activator by ras in intact cells and in a cell-free system J Biol Chem 1992 267 20346-20351
18 Montcouquiol M and Corwin JT Intracellular signals that control cell proliferation in mammalian balance epithelia key roles for phosphatidylinositol-3 kinase mammalian target of rapamycin and S6 kinases in preference to calcium protein kinase C and mitogen-activated protein kinase J Neurosci 2001 21 570-580
19 Juliano RL Signal transduction by cell adhesion receptors and the cytoskeleton functions of integrins cadherins selectins and immunoglobulin-superfamily members Annu Rev Pharmacol Toxicol 2002 42 283-323
20 Rosales JL et al GTP-dependent secretion from neutrophils is regulated by Cdk5 J Biol Chem 2004 279 53932-53936
21 Shibuya M Structure and function of VEGFVEGF-receptor system involved in angiogenesis Cell Struct Funct 2001 26 25-35
22 Gafni J et al Inhibition of Calpain Cleavage of Huntingtin Reduces Toxicity accumulation of calpaincaspase fragments in the nucleus J Biol Chem 2004 279 20211-20220
Huntingtons Disease Antibodies Proteins and Peptides
Huntingtons Disease
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
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1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Internet sigma-aldrichcom
10
Huntingtons Disease Signaling For this and related interactive pathways see sigmacomhdsig
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 11
Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Gene Transcription
Cell SurvivalNeurodegeneration
Caspase 8Caspase 8
Caspase 37
Caspase 12
Calpain
Ca2+
Ca2+
GRIN2BCa2+ ChannelN-type
GRM15
Caspas
Ca2+
Ca2+
DecreasedMitochondrial
Membrane PotentialExcitotoxic Death Apoptosis
Neurodegeneration
HD
Bax
HD
NeuronalIntranuclear
Inclusion
Cytochrome c
CaCC 2++
Ca
Baax
HD
Gβ
Gγ
se
HIP 1
HD
se 12
HIP
HD
HIPPI
Bcl-xI
HIP 11HIP 1
Bcl-xIB l I
Altered in HD
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HD NH2-TerminalFragment Aggregates
HDFragment
HD
HD
HD
HD
HD
HIPPIHIPPI
HD
1
Apaf 1
CCaspas
ochrome c
CCaspas
Apaf 1
Calpain
CDK5
p35
SGK AKT
PDK1
PIP3IP3
GαqPLCβ
PSD-95
PKC DAG
PI3K
IGF1R
IGF-1
PIP2PIP2
HAP1
Caspase
mTOR
Ca2+
PIP33PIP22
HDFragment
HDCse
HD Fragment
Neuro
ragment
P
HD
p35
HD P
HD
HD
HD
Ca2+
PIP2
SGK
HAP1
HD NH2-TerminalFragment Aggggregates
DAG
D
HD
HD
HDFragment
DNAJC5
HDFragment
HD
HD
DNAJC5DSTX1A
HD
Ca2+
Glutamate
Ca2+
GR
GlutamateCa2+
a2+ Channel
PHD
NCOR
TGM2CBP
HD Fragment
CB
NeuI t
SH3GL3
uclearsion
nuus
IntranInclu
p53
sionusInclu
SP1
p53
SIN3A
SP1
CA150BP
TGBP
GMGGMM2M2
ntennten SIN3A
CA150TBP
NCOOR
SH3GGL33
HD FraHD Fraaaaaagmagmmeme
TAFII130
CBP HDFragment
TAFII130TA H0AFII130
TFIIDCBPFra
HDntagmen
TFIID
TBP
Gene Transcription
EnkaphalinEnkaphalin
HIP 1HIP 1
HIPPI
HIP 1
HD
HIP 1
HDHDDDHD
CREB
CRE
CREB
CRE
Autophagy
Polymerase II
Cytochrome c
Mitochondria
SR
mTOR
Arfaptin 2ArfaptinP
IP33
HDMisfolded
HP14
Endocytosis
HD
HP14HP14HP14
HD
Impaired Exocytosis
SNARE RPH3A
Reduced in HD Reduced in HD
SynapticVesicle
1
HDD SH3GL3 HDPACSIN1 HPACCSIN1
n 2P
HD
E
d in HD
RPH3A
Reduced in HDSTX1A
HD NH2-TerminalFragment Aggregates
Growth andDierentiation
bull Neurite Outgrowth bull Survival bull Proliferation bull Differentiation
EGF
HD
HD
RASGAP
hRas
HD
HD
HDJNK1
NGF
PI3K
hRas
ERK 12AKT1
HD
EGF
EGFR EGFR EGFR EGFR TRKA TRKA
HD
HD
hRas
HD
SOS
GBR2
SOS
GBR2 SHC HD hRas
GRB2
SOSSSHC
RASGAP
HD AA
HD HAP1
HD
HDMLK2
MLK2
MLK2
MLK2
NeuroD
MKK 47
JNK2
JNK2
p53C-Jun
P1
Gene Transcription
NeuronalDevelopment and
SurvivalApoptosis
HD
MLK2
MLK2
HD HAP
HD
MLK2
NeuroD
p53p53C-Junun
P
HDHD
P
Gene Transcription
BDNF
Cell Death
REST
RESTREST
HD
RESTR
HD
R
HD
RCOR HDAC
SIN3A
RRCOR HDAC
REST
REST
REST
REST
REST HD
Polymerase II
BDNF
NRSE
PerinuclearInclusions
Cell Death
Proteasome
Protein Degradation
dHD
Misfolded
Ub
Ub
Hsp70
Hsp40
HD
HDMisfolded
UBE2S
SH3GL3
Hsp40
HDFragment
HDMisfolded
HIP1
DCTN1
HIP1
TN1DCT
SNCADNM
GLS
SDH
Hsp40Hsp40HH
ATP5
DCTN1
Hsp40
HAP1
DYNC1I2
1DCTN1HAP1
DYNC1I2DYNC1I2
HD
HAP1HAP1
HD
Axonal TransportAlong Microtubules
NeurotoxicityDeath
MitochondrialTracking
MitochondrialDysfunction
HDHD
HIP1 CLTC
Hsp40
AP2
HHIP1 CLTC
AP2
HD
Clathrin-coated
Vesicles
Clathrin-coated
Vesicles
Clathrin-t d
BDNF
Vesiccles
BDNF
11
0Hsp40
siclesVes
TGM2
MisfDDHHHDHDdedfoold
TN1DCT
SNCASDH
Ub
D
rinuclearclusions
LSGLSUb
SH3GGL3HHHD
Hsp70CTSD
DHD
Dment
DNMDNM
t
MM S
DHHgFragHDm
DD
HHHD
Hsp70CTSD
Microtubules
Cytoplasm
Extracellular Space
Nucleus
ITPR1
HD
P PP P
P P
Caspase 9
HD
Appaf
Casppase 9p
CPLX2SNARE
Reduced
CPPLX2
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
scTrans
oa
micro
d
pp
oac
id
p
r ctor
Pepti
Phos
Kina
gaepeucece
Ph
LigdeNuRe
el
p
cca
aa
scTranscca
lTranslRegulaRegul
TranscRegulTrans
t
Mutat
t
Transp
Othe
P Ubiquitinated Protein
Ub
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Internet sigma-aldrichcom
12
Antibodies for Huntingtons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoMonoclonal Anti-BDNF mouse 1B10 BDNF human human ELISA (i)
WB- SAB1402127-100UG
Monoclonal Anti-CREBBP mouse 2B6 CREBBP human human ELISA (c)ELISA (i)
WB
- SAB1403694-100UG
Anti-DLGAP2 rabbit - DLGAP2 human human IHC (p)PAWB
- HPA030320-100UL
Anti-EP300 rabbit - EP300 human human IF (i)IHC (p)
PA
- HPA003128-100UL
Anti-Glutamate Receptor NMDAR1 (NR1)
rabbit - GRIN1 humanGrin1 rat
Grin1 mouse
humanmouse
rat
WB - G8913-2ML
Anti-HAP1 (C-terminal) rabbit - HAP1 human human WB - SAB4200293-200UL
Anti-HIP1 rabbit - HIP1 human human IF (i)IHC (p)
PAWB
- HPA013606-100UL
Anti-HIP14 rabbit - Zdhhc17 mouseZDHHC17 human
bovinecaninehumanmouse
rat
WB H7414-25ULH7414-200UL
Monoclonal Anti-Histone Deacetylase 1 (HDAC1)
mouse HDAC1-21 Hdac1 mouseHDAC1 human
humanmouse
ARRELISA (i)
IPWB
- H6287-200UL
Monoclonal Anti-Histone Deacetylase 2 (HDAC2)
mouse HDAC2-62 HDAC2 humanHdac2 mouse
Hdac2 rat
bovinecaninechickenhumanmouse
rat
ARRELISA (i)
IHCIP
WB
- H2663-200UL
Monoclonal Anti-Histone Deacetylase 4 (HDAC4)
mouse HDAC4-144 Hdac4 ratHDAC4 humanHdac4 mouse
humanmouse
rat
ICCIP
WB
- H0163-200UL
Monoclonal Anti-Histone Deacetylase 5 (HDAC5)
mouse HDAC5-35 HDAC5 humanHdac5 mouse
Hdac5 rat
humanmouse
rat
ARRELISA (i)
ICCIP
WB
- H4538-200UL
Anti-MAP3K10 (867-880) rabbit - MAP3K10 human human WB - M6571-200UL
Anti-MAPK9 (276-290) rabbit - MAPK9 human human WB - M7573-200UL
Anti-NeuroD1 rabbit - NEUROD1 humanNeurod1 rat
Neurod1 mouse
humanmouse
rat
WB - N3663-25ULN3663-200UL
Monoclonal Anti-Polyglutamines mouse 3B5H10 HTT human human ICCIP
WB
P1874-200UL
Anti-REST rabbit - REST human human IF (i)IHC (p)
PA
- HPA006079-100UL
Anti-Sin3A C-Terminal rabbit - Sin3a ratSIN3A humanSin3a mouse
human ARRIP
WB
- S6695-200UL
Monoclonal Anti-TBP mouse 58C9 Tbp Drosophila melanogasterTBP human
Drosophila melanogasterSf9 cell line
humanyeast
IPWB
- T1827-25ULT1827-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
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Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
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Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 13
Immunofluorescence of HUVEC cells using MAP3K10 (867-880) (RB) Cat No M6571 Yale HTCB IF procedure used
Anti-REST Cat No HPA006079 Immunofluorescent staining of human cell line U-2 OS
Proteins amp Peptides for Huntingtonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No
8 Bcl-xL Active human BCL2L1 is a member of the BCL2 apoptotic regulators that interacts with the voltage-dependent anion channel VDAC The long isoform inhibits apoptosis whereas the short isoform promotes cell death Human Bcl-xL (amino-acids 1-212) GenBank Accession No Z23115 with C-terminal His tag MW = 28 kDa expressed in an E coli expression system
BCL2L1 ge90 SDS-PAGE SRP0187-100UG
8 BDNF human BDNF is a member of the NGF family of neurotrophic growth factors that supports neuron proliferation and survival Expression is reduced in both Huntingtons and Alzheimers disease
BDNF ge98 HPLCge98 SDS-PAGE
SRP3014-10UG
8 Calpain 1 human Cytosolic protease with involvement in cytoskeletal remodeling autophagy and apoptosis as an upstream regulator
CAPN1 ge95 SDS-PAGE C6108-100UG
8 CBP (1319-1710) GST tagged human
CREB-binding protein (CREBBP) binds specifically to phosphorylated CREB enhancing cAMP-responsive transcriptional activity 1319-1710 contains the catalytic domain for lysine acetylation activity
CREBBP ge70 SDS-PAGE SRP5173-50UG
8 KAT3A (518-1207) GST tagged human
KAT3A (CREBBP) mediates coactivation of many transcription factors It couples chromatin remodeling to transcription factor recognition via its intrinsic acetyltransferase activity playing a key role in development and growth control
CREBBP ge70 SDS-PAGE SRP5219-20UG
8 CoREST human Human recombinant CoREST GenBank Accession No NM_015156 amino acids 305-end with N-terminal His tag MW = 20 kDa expressed in E coli expression system
RCOR1 ge60 SDS-PAGE SRP0124-100UG
8 HDAC-1 human Useful for the study of enzyme kinetics and screening inhibitors Human HDAC1 GenBank Accession No NM_004964 full length with C-terminal HIS-DDDDK tag (FLAGreg) and C-terminal His-tag MW = 56 kDa expressed in baculovirus expression system
HDAC1 ge50 SDS-PAGE SRP0100-50UG
8 HDAC-2 His tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal His tag MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge95 SDS-PAGE SRP0102-50UG
8 HDAC-2 FLAG tag human Human HDAC2 GenBank Accession No Q92769 full length with C-terminal DDDDK tag (FLAGreg) MW = 60 kDa expressed in baculovirus expression system
HDAC2 ge50 SDS-PAGE SRP0103-50UG
8 HDAC-4 human Human HDAC4 GenBank Accession No NM_006037 amino acids 627-1085 with N-terminal ST tag MW = 752 kDa expressed in baculovirus expression system
HDAC4 ge50 SDS-PAGE SRP0105-2UG
8 HDAC-5 full length human Human HDAC5 GenBank Accession No NM_001015053 full length with N-terminal ST tag MW = 150 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0107-5UG
8 HDAC-5 human Human HDAC5 catalytic domain GenBank Accession No NM_001015053 amino acid 657-1123 with C-terminal His tag MW = 51 kDa expressed in baculovirus expression system
HDAC5 ge80 SDS-PAGE SRP0106-5UG
IGF-I from rat IGF-I is a member of a family of polypeptide growth factors that mediate growth and development IGF-I has been linked to neuroplasticity and hippocampal neurogenesis IGF-I (Insulin-like Growth Factor-I) is a polypeptide growth factor that stimulates the proliferation of a wide range of cell types including muscle bone and cartilage tissue Rat IGF-I is a 769 kDa protein containing 70 amino acid residues
Igf1 ge95 HPLCge95 SDS-PAGE
SRP4121-20UG
14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
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Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
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ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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14
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Insulin-like Growth Factor-I
humanIGF-I also known as somatomedin C mediates the growth-promoting activity of GH (growth hormone) IGF-I induces endothelial cell migration and is involved in the regulation of angiogenesis IGF-I exerts its actions through the IGF-I receptor
IGF1 ge97 SDS-PAGE or HPLC
I3769-50UG
Insulin-like Growth Factor-I from mouse
Potent mitogenic growth factor that mediates the growth-promoting activities of growth hormone postnatally Mouse and human IGF-I share 97 sequence identity
Igf1 gt97 SDS-PAGE I8779-50UG
Insulin-like Growth Factor-I (E3R) human
IGF1(3R) contains the amino acid substitution E3R which decreases binding to IGF-binding proteins IGF1(3R) therefore is much more potent than the native IFG1
IGF1 gt95 HPLC I2656-25UG
IGF1R (960-end) active His tagged human
IGF1R is a transmembrane tyrosine kinase receptor that mediates the effects of IGF1 and thus plays an important role in growth and development including cerebellar development and hippocampal neuronal plasticity
IGF1R ge70 SDS-PAGE I0786-10UG
Insulin-like Growth Factor-I Receptor human
Binds IGF-I with high affinity IGF-II with lower affinity and insulin with weak affinity
IGF1R ge95 SDS-PAGE I4657-50UG
8 JNK2 active GST tagged human
JNK2 (MAPK9) acts as an integration point involved in a wide variety of cellular processes such as proliferation differentiation transcription regulation and development including regulation of regional specific apoptosis during early brain development
MAPK9 ge70 SDS-PAGE SRP5042-10UG
c-Jun human Substrate for SAPK1JNK2 JUN 40-50 SDS-PAGE C5859-100UG
8 p300 human EP300 and CREBBP are highly related transcriptional coactivators mediating cAMP gene regulation EP300 is known to acetylate many transcription factors including p53 E2F TFIIE and TFIIF
EP300 ge70 SDS-PAGE SRP2079-4UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6249-50UG
p53 human p53 gene is highly conserved and expressed in normal tissues It is the most commonly mutated gene in human cancer and more then 500 gene mutations have been described in various types of malignancies hematologic as well as solid tumors Intact p53 function is essential for the maintenance of the non-tumorogenic phenotype of cells Thus p53 plays a vital role in suppressing the development of cancer
TP53 ge90 SDS-PAGE P6374-20UG
p53 Mutant human p53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth replication and apoptosis Mutation involving p53 is found in a wide variety of malignant tumors including breast ovarian bladder colon lung and melanoma
TP53 ge90 HPLCge90 SDS-PAGE
SRP4832-5UG
PDK1 Active human PDK1 plays a regulatory role in glucose and carbohydrate metabolism It functions through the PI3K signaling cascade upstream of AKT1 as well as in other pathways involving proliferation survival and cell migration
PDPK1 ge75 SDS-PAGE P7498-5UG
PDK1 active His tagged human
PDK1 plays a key regulatory role in the homeostasis of carbohydrate fuels It activates protein kinase B (PKB) which in turn inactivates GSK3 and may potentiate the effects of IGF1
PDPK1 ge70 SDS-PAGE K3393-10UG
RACK1 human RACK1 is an intracellular receptor protein that binds activated members of the protein kinase C family
GNB2L1 gt90 SDS-PAGE R4780-50UG
8 TATA box binding protein GST tagged human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge70 SDS-PAGE SRP2106-10UG
8 TBP (TATA box binding protein) human
TBP is the core protein of the transcription factor IID complex the multiprotein DNA-binding factor that coordinates activities necessary for initiation of transcription by RNA polymerase II
TBP ge85 SDS-PAGE SRP2003-10UG
To view additional products for Huntingtons Disease Research visit sigmacomhunt
Antibody catalog numbers beginning with HPA are Prestige Antibodiesreg powered by Atlas Antibodies
Proteins amp Peptides for Huntingtonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 15biomolecules
BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
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BiocharacterizedHighly characterized neuropeptides from Sigmareg
Sigma Life Science is the leading provider of peptides for your neuroscience research Our highly characterized neuropeptides offer the high purity and specificity that your research demands
with all of our specifications and analysis available on the web
Visit sigmacomneuropeptides to browse β-amyloids neurotransmitters vasoactives and more
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLCregistered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
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SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
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16
Parkinsons disease (PD) is the second most common neurodegenerative disorder after Alzheimers disease affecting more than 6 million people worldwide PD is a slowly progressing motor system neurodegeneration characterized by akinesia rigidity and resting tremor Neuropathologically PD is characterized by loss of dopaminergic cell bodies in the substantia nigra resulting in a reduced supply of dopamine to the basal ganglia The high metabolic rate of the substantia nigra combined with high content of oxidizable species and iron high levels of reactive oxygen species (ROS) and low level of antioxidants all serve to initiate and propagate apoptosis of the dopaminergic neurons
Mutations in the α-synuclein gene (SNCA) occur in familial cases of Parkinsons disease pointing to a role for this gene in PD α-Synuclein can form protein aggregates with additional cytoskeletal proteins including synaptophysin (SYP) and Tau (MAPT) which are believed to lead to the pathogenesis of Lewy body formation
Synphilin (SNCAIP) interacts with α-synuclein in neuronal tissue and is thought to play a role in the formation of cytoplasmic inclusions and neurodegeneration A mutation in this gene has been associated with Parkinsons disease
Mutations in the LRRK2 (PARK8) gene are found in about 5-6 percent of all familial cases as well as 2 percent of cases with no known cause Interestingly this mutation can cause early-onset Parkinsons in families from diverse ethnic backgrounds in a form that is
identical in clinical symptoms to late-onset Parkinsons LRRK2 encodes a protein that is part of a larger multidomain protein with characteristic GTPase and kinase domains LRRK2s substrates its binding partners and its regulators have yet to be confirmed or clarified and consequently its role in normal physiological functions in the cell and in disease are still largely unknown
Loss of function mutations in the Parkin (PARK2) PINK1 (PARK6) and PARK7 (DJ-1) genes resulting in functionally inactive proteins underlie common forms of autosomal-recessive PD Patients with loss-
of-function Parkin mutations account for an estimated 40-50 percent of all familial early-onset cases of PD whereas mutations in PINK1 and PARK7 are less common Several studies have demonstrated that products of all three recessive genes preserve mitochondrial functions protect against reactive oxygen species or play a role in protein degradation pathways Normally Parkin tags proteins with ubiquitin for degradation via the proteasome Mutations in the Parkin gene lead to a loss of this activity DJ-1 is a molecular chaperone involved in protein folding as well as in
Parkinsons Disease Antibodies Proteins and Peptides
Parkinsons Disease
SYPH1
p38MAPKJNK1
Synuclein-α
Cytoplasm
PAELR
UCHL1
PARK7 PARK3
PARK4
Lewy BodyFormation
Death ofDopaminergic
Neurons
Parkinsonrsquos Disease
Accumulation of PAELR in the ER
Inhibition ofDopamine Release
CytoplasmicAccumulation of
Dopamine
SYPH1 PAELR
PARK4
PARK3PARK7
UbSynuclein-αα
Ub Ub
Parkin Ub
P
Production ofReactive Oxygen
Species
Cytochrome c
Caspase 9
Caspase 3
O-glycosylgroup
UbiquitinationPathway
SEPT5Ub
Accumulation of O-glycosylated
Synuclein-αAccumulation of
SYPH1
Chemical Drug orToxicant
Cytokine or Growth Factor
Enzyme
G-proteinCoupledReceptor
Group orComplex
Ion Channel
Kinase
Phosphorylated Protein
Ligand-dependent NuclearReceptor
microRNA
Peptidase
Phosphatase
TranscriptionRegulator
TranslationRegulator
TransmembraneReceptor
Transporter
Other
Key
Note ldquoActs Onrdquo and ldquoInhibitsrdquo edge may also include a binding event
Direct Interaction
Indirect Interaction
A B Bindings Only
A B Inhibits
A B Inhibits and Acts On
A B Leads To
A B Translocates To
A B Reaction
A B
A B
Enzyme Catalysis Reaction
Disruption
Mutated Gene
A B Acts On
cal ccrrorn
nhh
microRNA
e
tt
rrrnnn
nnnhhhhh
se
at
rrt
e orFFactor
Peptidas
Phospha
e
phi
d-nd
earpto
Prote
Kinase
PhospP
LigandepenNucleRecep
ex
annel
Receptor
t
tr
or
m
m
teet
p le
e
ee
ex Transmem
eeinedtoor
orex
ptpro
oTranslatiorRegulato
Transmem
eein
po
T l ti
TranscripRegulato
eeee
ex Transmem
-dror Mutated
-dro
-dentr
Transport
Other
M d
P Ubiquitinated Protein
Ub
Parkinsons Signaling For this and related interactive pathways see sigmacompdsig
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 17
other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
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Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
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other functions It is found in the cytosol the mitochondrial matrix and intermembrane space It regulates redox-dependent signaling pathways and acts as a regulator of antioxidant gene expression
UCHL1 (PARK5) is a member of the ubiquitin-C-terminal hydrolases Expression of UCHL1 is highly specific to neurons and to cells of the diffuse neuroendocrine system and their tumors A point mutation I93M in this protein is implicated as the cause of PD Furthermore a polymorphism S18Y in this gene has been found to be associated with a reduced risk for Parkinsons disease UCH-L1 is also associated with the Alzheimers disease
The ATP13A2 (PARK9) gene encodes a member of the P5 subfamily of ATPases which transports inorganic cations as well as other substrates Mutations in this gene are associated with Kufor-Rakeb syndrome (KRS) also referred to as Parkinson disease 9 and in juvenile forms of PD
Additional genes including GIGYF2 (PARK11) HTRA2 (OMI PARK13) PLA2G6 (PARK14) FBXO7 (PARK15) STUB1 (CHIP) and RNF19A (Dorfin) are thought to be implicated in PD
With its complex etiology and impact on millions Parkinsons Disease continues to be the subject of intensive research effort
References1 Pawlyk AC et al Novel monoclonal antibodies
demonstrate biochemical variation of brain parkin with age J Biol Chem 2003 278 48120-48128
2 Kitada T et al Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism Nature 1998 392 605-608
3 Maroteaux L and Scheller RH The rat brain synucleins family of proteins transiently associated with neuronal membrane Mol Brain Res 1991 11 335-343
4 Ueda K et al Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease Proc Natl Acad Sci USA 1993 90 11282-11286
5 Kurihara LJ et al Loss of Uch-L1 and Uch-L3 leads to neurodegeneration posterior paralysis and dysphagia Hum Mol Genet 2001 10 1963-1970
6 Leroy E et al The ubiquitin pathway in Parkinsons disease Nature 1998 395 451-452
7 Levecque C et al No genetic association of the ubiquitin carboxy-terminal hydrolase-L1 gene S18Y polymorphism with familial Parkinsons disease J Neural Transm 2001 108 979-984
8 Liu Y et al The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinsons disease susceptibility Cell 2002 111 209-218
9 Lee G et al Synphilin-1 degradation by the ubiquitin-proteasome pathway and effects on cell survival J Neurochem 2002 83 346-352
10 Nagano Y et al Siah-1 facilitates ubiquitination and degradation of synphilin-1 J Biol Chem 2003 278 51504-51514
11 Liani E Ubiquitylation of synphilin-1 and alpha-synu-clein by SIAH and its presence in cellular inclusions and Lewy bodies imply a role in Parkinsons disease Proc Natl Acad Sci USA 2004 101 5500-5505
12 Arai R et al Differential subcellular location of mitochondria in rat serotonergic neurons depends on the presence and the absence of monoamine oxidase type B Neurosci 2002 114 825-835
13 Fernandez HH and Chen JJ Monamine oxidase inhibitors current and emerging agents for Parkinson disease Clin Neuropharmacol 2007 30 150-168
14 Hishikawa N et al Dorfin localizes to the ubiquitylated inclusions in Parkinsons disease dementia with Lewy bodies multiple system atrophy and amyotrophic lateral sclerosis Am J Pathol 2003 163 609-619
15 Ito T et al Dorfin localizes to Lewy bodies and ubiquitylates synphilin-1 J Biol Chem 2003 278 29106-29114
16 Paisan-Ruiz C et al Cloning of the gene containing mutations that cause PARK8-linked Parkinsons disease Neuron 2004 44 595-600
17 West AB et al Parkinsons disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity Proc Natl Acad Sci USA 2005 102 16842-16847
18 Cookson MR et al The roles of kinases in familial Parkinsons disease J Neurosci 2007 27 11865-11868
19 Valente EM et al Hereditary early-onset Parkinsons disease caused by mutations in PINK1 Science 2004 304 1158-1160
20 Beilina A et al Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability Proc Natl Acad Sci USA 2005 102 5703-5708
21 Park J et al Mitochondrial dysfunction in Drosophila PINK1 mutants is complemented by parkin Nature 2006 441 1157-1161
22 Torres GE et al Plasma membrane monoamine transporters structure regulation and function Nature Rev Neurosci 2003 4 13-25
23 Ciliax BJ et al Immunocytochemical localization of the dopamine transporter in human brain J Comp Neurol 1999 409 38-56
24 Gandhi S et al PINK1 protein in normal human brain and Parkinsons disease Brain 2006 129 1720-1731
25 Ramirez A et al Hereditary parkinsonism with dementia is caused by mutations in ATP13A2 encoding a lysosomal type 5 P-type ATPase Nature Genet 2006 38 1184-1191
26 Wei J et al Enhanced lysosomal pathology caused by beta-synuclein mutants linked to dementia with Lewy bodies J Biol Chem 2007 282 28904-28914
27 Ning YP et al PARK9-linked parkinsonism in eastern Asia mutation detection in ATP13A2 and clinical phenotype Neurol 2008 70 1491-1493
28 Hod Y Differential control of apoptosis by DJ-1 in prostate benign and cancer cells J Cell Biochem 2004 92 1221-1233
29 Bonifati V et al Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism Science 2003 299 256-259
30 Faccio L et al Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia J Biol Chem 2000 275 2581-2588
31 Strauss KM et al Loss of function mutations in the gene encoding OmiHtrA2 in Parkinsons disease Hum Mol Genet 2005 14 2099-2111
32 Park HJ et al Beta-amyloid precursor protein is a direct cleavage target of HtrA2 serine protease Implications for the physiological function of HtrA2 in the mitochondria J Biol Chem 2006 281 34277-34287
33 Morgan NV et al PLA2G6 encoding a phospholipase A2 is mutated in neurodegenerative disorders with high brain iron Nat Genet 2006 38 752-754
34 Gregory A et al Neurodegeneration associated with genetic defects in phospholipase A(2) Neurol 2008 71 1402-1409
35 Schaeffer EL and Gattaz WF Cholinergic and glutamatergic alterations beginning at the early stages of Alzheimer disease participation of the phospholipase A2 enzyme Psychopharmacol 2008 198 1-27
36 Petrucelli L et al CHIP and Hsp70 regulate tau ubiquitination degradation and aggregation Hum Mol Genet 2004 13 703-714
37 Shin Y et al The co-chaperone carboxyl terminus of Hsp70-interacting protein (CHIP) mediates alpha-synuclein degradation decisions between proteasomal and lysosomal pathways J Biol Chem 2005 280 23727-23734
38 Miller VM et al CHIP suppresses polyglutamine aggregation and toxicity in vitro and in vivo J Neurosci 2005 25 9152-9161
18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
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18
Antibodies for Parkinsons ResearchProduct Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-ATP13A2 (C-terminal region) rabbit - ATP13A2 human human
mouseWB - A3361-25UL
A3361-200UL
Anti-Caspase 3 Active rabbit - Casp3 mouseCASP3 human
Casp3 rat
bovinecaninehumanmouse
pigrat
ARRIF (i)WB
- C8487-200UL
Anti-Caspase 9 rabbit - CASP9 humanCasp9 rat
humanrat
ARRIHC (p)
IPWB
- C7729-2ML
Anti-CHIP (N-terminal) rabbit - Stub1 mouseK08D1011 ratSTUB1 human
humanmouse
rat
WB - C9118-25ULC9118-200UL
Anti-Cytochrome c sheep - CYCS humanCycs rat
caninehumanrabbit
rat
IF (i)IHC (p)
WB
- C9616-200UL
Monoclonal Anti-DOPA Decarboxylase (DDC)
mouse DDC-109 DDC humanDdc rat
bovinecanine
guinea pighumanmonkey
rabbitrat
sheep
ARRELISA (i)
ICCIP
WB
D0180-2MLD0180-5ML
Anti-Dorfin (N-terminal) rabbit - Rnf19a mouseRNF19A human
humanmouse
rat (predicted)
WB D0319-25ULD0319-200UL
Anti-HtrA2 rabbit - Htra2 ratHTRA2 human
humanrat
WB H7290-25ULH7290-200UL
Monoclonal Anti-LRRK2 mouse PROK57 LRRK2 humanLrrk2 mouse
humanmouse
ELISA (i)ICCWB
L3044-25ULL3044-200UL
Anti-LRRK2 (C-terminal region) rabbit - Lrrk2 mouseLRRK2 human
Lrrk2 rat
humanmouse
rat
WB L9918-25ULL9918-200UL
Anti-MAPK11 (306-320) rabbit - MAPK11 human human WB - M1322-200UL
Anti-MAPK13 (333-345) rabbit - MAPK13 human human WB - M1572-200UL
Monoclonal Anti-p38 MAP Kinase Activated (Diphosphorylated p38)
mouse P38-TY Mapk14 ratMapk14 mouseMAPK14 human
humanmouse
rat
ARRELISA (i)
ICCWB
- M8177-2ML
Anti-PARK7 rabbit - PARK7 human human IF (i)IHC (p)
PAWB
- HPA004190-100UL
Monoclonal Anti-Parkin mouse PRK8 Park2 ratPARK2 humanPark2 mouse
hamsterhumanmouse
rat
ARRWB
P6248-200UL
Anti-Phospholipase A2 (iPLA2) rabbit - pla2g6 ratpla2g6 mouse
PLA2G6 human
humanmouse
rat
WB - SAB4200129-25ULSAB4200129-200UL
Anti-PINK1 rabbit - Pink1 ratPink1 mousePINK1 human
humanmouse (predicted)
rat (predicted)
WB P0076-25ULP0076-200UL
Anti-SEPT5 (1-14) rabbit - SEPT5 human human IF (i)WB
- SAB1100586-200UL
Anti-SEPT5 (316-330) rabbit - SEPT5 human human IF (i)WB
- SAB1100587-200UL
Anti-Synphilin-1 rabbit - Sncaip ratSncaip mouseSNCAIP human
human WB S5946-200UL
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052 Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice DevelopmentCustom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrichcomsafetycenter
Sigma-Aldrichreg Worldwide Offices
NYD77157-510122
1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
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Internet sigma-aldrichcom
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 19
Product Name Host Clone No Gene Symbol Species Reactivity Application Array Antibody Cat NoAnti-Synphilin-1 (C-terminal) rabbit - SNCAIP human
Sncaip ratSncaip mouse
human WB S6071-200UL
Anti-α-Synuclein rabbit - Snca ratSNCA human
humanrat
ARRIHC (p)
WB
S3062-2ML
Anti-Ubiquitin C-terminal Hydrolase L1 (RA-15)
rabbit - Uchl1 mouseUchl1 rat
UCHL1 human
humanmouse
rat
ARRWBWB
U5258-200UL
Proteins amp Peptides for Parkinsonprimes Research Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No Caspase 3 human Member of the CED-3 subfamily of caspases and responsible for the cleavage
of many key proteins such as the nuclear enzyme poly(ADP-ribose) polymerase (PARP) the inhibitor of caspase-activated deoxyribonuclease (ICAD) and gelsolin a protein involved in apoptosis regulation Human recombinant C-terminal histidine tagged caspase 3 is a fully active protein consisting of 17 kDa and 135 kDa subunits the 135 kDa subunit contains the histidine tag
CASP3 ge90 SDS-PAGE C1224-10UG
Caspase 9 human Activation of caspase-9 (CASP9) through apoptotic stimuli initiates the caspase cascade Caspases have been implicated in many disorders including cancer inflammatory disease neurodegenerative diseases stroke and myocardial infarction
CASP9 ge90 SDS-PAGE C8726-25UG
Cytochrome c from human heart
Cytochrome c is a small heme protein that is a mobile electron carrier in the mitochondrial electron transport chain It is also an initiator of apoptosis
CYCSL1CYCS
ge95 SDS-PAGE C3483-10UG
JNK1 active GST tagged from mouse
JNK1 is a member of the MAP kinase family that plays a role in the induction of apoptosis Disruption of MAP kinase signaling functionality is associated with disease states including inflammation cancer and neurodegenerative disease
Mapk8 ge70 SDS-PAGE J2455-10UG
p38α active GST tagged human
MAPK14 is a member of the p38 MAPK family members of which are activated by various environmental stresses and proinflammatory cytokines Suggested roles of this kinase include involvement in stress related transcription cell cycle regulation and genotoxic stress response
MAPK14 ge70 SDS-PAGE A4861-10UG
p38β active GST tagged human
p38β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress The p38β is activated through its phosphorylation by MAP kinase kinases (MKKs) preferably by MKK6 Transcription factor ATF2CREB2 has been shown to be a substrate of this kinaseAlternatively spliced transcript variants encoding the same protein have been observed
MAPK11 ge70 SDS-PAGE B4437-10UG
Immunofluorescence of HUVEC cells using MAPK11 (306-320) (RB) Cat No M1322 Yale HTCB IF procedure used
Anti-PARK7 Cat No HPA004190 Immunofluorescent staining of human cell line U-2 OS
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052 Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice DevelopmentCustom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrichcomsafetycenter
Sigma-Aldrichreg Worldwide Offices
NYD77157-510122
1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
sigma-aldrichcom
Enabling Science to Improve the Quality of Life
ArgentinaFree Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698
AustraliaFree Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500
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BrazilFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895
CanadaFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292
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Czech RepublicTel (+420) 246 003 200 Fax (+420) 246 003 291
DenmarkTel (+45) 43 56 59 00 Fax (+45) 43 56 59 05
FinlandTel (+358) 9 350 9250 Fax (+358) 9 350 92555
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MexicoFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel (+52) 722 276 1600 Fax (+52) 722 276 1601
The Netherlands Tel (+31) 78 620 5411 Fax (+31) 78 620 5421New ZealandFree Tel 0800 936 666 Free Fax 0800 937 777 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500
NorwayTel (+47) 23 17 60 00 Fax (+47) 23 17 60 10
PolandTel (+48) 61 829 01 00 Fax (+48) 61 829 01 20
PortugalFree Tel 800 202 180 Free Fax 800 202 178 Tel (+351) 21 924 2555 Fax (+351) 21 924 2610
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SwedenTel (+46) 8 742 4200 Fax (+46) 8 742 4243
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ThailandTel (+66) 2 126 8141 Fax (+66) 2 126 8080
United KingdomFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313
United StatesToll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757
VietnamTel (+84) 8 3516 2810 Fax (+84) 8 6258 4238
Internet sigma-aldrichcom
20
Product Name BiochemicalPhysiological Actions Gene Symbol Purity Cat No p38γ active
GST tagged humanp38γ is a member of the p38 MAPK family which is activated in response to stress p38γ gene was mapped to 22q133 and functions as a signal transducer during differentiation of myoblasts to myotubes Enforced localization of p38γ in the nucleus or cytoplasm markedly attenuates the ability of the kinase to induce cell cycle arrest in fibroblasts p38γ increases basal glucose uptake and decreases DNP- and contraction-stimulated glucose uptake partially by affecting levels of glucose transporter expression in skeletal muscle
MAPK12 ge70 SDS-PAGE G8546-10UG
p38δ active GST tagged human
p38δ (SAPK4) is a member of the p38 MAPK family and is activated by chemical and environmental stresses as well as by proinflammatory cytokines p38δ has a TGY dual phosphorylation motif and is activated in response to cellular stresses and proinflammatory cytokines MAP kinase kinases 3 and 6 can phosphorylate and activate this kinase Transcription factor ATF2 and microtubule dynamics regulator stathmin have been shown to be the substrates of this kinase
MAPK13 ge70 SDS-PAGE D7444-10UG
Parkin N-Terminal Peptide Used as marker for degradation of parkin PARK2 ge60 HPLC P2615-1MG
α-Synuclein human 140-amino acid protein (apparent molecular mass 19-20 kDa) that induces polymerization of tubulin into microtubules and functions in the modulation of dopamine
SNCA ge90 SDS-PAGE S7820-500UG
α-Synuclein A30P human A point mutation in the α-synuclein gene Ala30-Pro (A30P) linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1196-500UG
α-Synuclein A53T human A point mutation in the α-synuclein gene A53T (Ala53-Thr) is linked to familial Parkinsons disease
SNCA ge90 SDS-PAGE S1071-500UG
α-Synuclein E46K human Deposition of α-synuclein as fibrillary tangles is a hallmark of certain neurodegenerative diseases including Parkinsons Among the familial mutations of α-synuclein E46K has the greatest potential to aggregate
SNCA ge90 SDS-PAGE S4447-500UG
8 UCHL1 His tagged human UCHL1 has ligase and hydrolase activities which play roles in proteasomal protein degradation a process critical for neuronal health Mutations in UCHL1 may be associated with Parkinsons disease and UCHL1 reverses the inhibition of CREB phosphorylation induced by Amyloid-β
UCHL1 ge70 SDS-PAGE SRP5149-50UG
To view additional products for Parkinsons Disease Research visit sigmacompark
Proteins amp Peptides for Parkinsonprimes Research continued
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052 Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice DevelopmentCustom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrichcomsafetycenter
Sigma-Aldrichreg Worldwide Offices
NYD77157-510122
1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
sigma-aldrichcom
Enabling Science to Improve the Quality of Life
ArgentinaFree Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698
AustraliaFree Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500
AustriaTel (+43) 1 605 81 10 Fax (+43) 1 605 81 20
BelgiumTel (+32) 3 899 13 01 Fax (+32) 3 899 13 11
BrazilFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895
CanadaFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292
ChileTel (+56) 2 495 7395 Fax (+56) 2 495 7396
Peoplersquos Republic of ChinaFree Tel 800 819 3336 Tel (+86) 21 6141 5566 Fax (+86) 21 6141 5567
Czech RepublicTel (+420) 246 003 200 Fax (+420) 246 003 291
DenmarkTel (+45) 43 56 59 00 Fax (+45) 43 56 59 05
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Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 21
Antibody microarrays are used for profiling complex protein samples and for assessing differential protein expression The Panorama Ab Microarray kits consisting of a series of arrays that cover various biological pathways such as Cell Signaling Gene Regulation and MAPKPKC pathways have already proven to be useful tools for such applications A new antibody array consisting of 224 neurobiology related antibodies has been developed Refer to Figure 1 for antibody distribution
Using this array we compared brain tissue of newborn and adult rats in order to study protein expression during rat brain development We identified several proteins that changed with age The outline of the experimental procedure is highlighted in Figure 2
Differential protein expression in rat models using the Panoramareg Neurobiology Array
Panoramareg Neurobiology Array
Developmental 40
BiopolarDepression 1
Parkinsons 13
Schizophrenia 9
Alzheimers 21
ALS 8Huntingtons 6
MS 2
Figure 1 ndash Distribution of neurospecific antibodies comprising the Panorama Neuorobiology Array
Step 2Label samples with Cy3Cy5
and mix
Step 3Incubate on the array
Step 4Scan the array
Sample BSample A
Step1Extract proteins
Figure 2 ndash Neurobiology Antibody Array Procedure
For a listing of antibodies on the array scan the QR code or visit sigmacomnbaa5
22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
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22
Expression of proteins were prepared from rat newborn (4 days) and adult (2 month) brains using ExtractionLabeling Buffer (as supplied in Panoramareg Neurobiology kit) and labeled with Cy3trade Equal amounts of labeled extracts (20 μgml) were incubated on the Neurobiology slides Proteins on left (indicated in green) demonstrate decline in development Proteins on right (indicated in
red ie Synaptopodin Ubiquitin C-terminal Hydolase L1 and alphabeta SNAP) are associated with increase with age Figure 3
Array results were confirmed by immunoblotting Equal amounts of protein extract (20 μg per lane) from rat adult (A) or newborn (N) brains were separated by SDS-PAGE and blotted onto nitrocellulose
membrane The proteins were probed with the monoclonal or polyclonal antibodies corresponding to the array and visualized using chemiluminescence
Further studies using the Panorama Neurobiology array have been performed identifying several additional proteins that changed with age For example PINK1 was shown to be elevated in adult versus newborn brains Further evaluation of the new Panorama Antibody Neurobiology Array was performed using brain tissue from the knockout rat model PARK2 -- The array has demonstrated that indeed this gene expression is being ablated in the knockout rats versus wild type siblings a result validated by immunoblotting (data not shown)
In conclusion the new Panorama Neurobiology array has shown to be a promising and useful tool for high-throughput screening (HTS) of protein level changes in neuronal development and neurodegenerative disease
Newborn Adult
AL S2CL (N-terminal region)
AL S2CL (N-terminal)
TAU
Synaptopodin
Ubiquitin C-terminalHydrolase L1
αβ SNAPHigh level
Low level
A N
A N
A N
A N
Figure 3 ndash Differential Expression between Newborn and Adult Rats
biomolecules
BioguaranteeSigmareg Life Science offers a collection of more than 50000 antibodies all 100 BioguaranteedFind the antibody you needsigmacomantibodyexplorer
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries Where bio begins is a trademark of Sigma-Aldrich Co LLC
Experimental results must be submitted via the Antibody Bioguarantee Form within 12 months of the date of purchase All required fi elds of the Antibody Bioguarantee Form must be completed Refunds and replacements contingent to claim review by technical service team Credit covers the cost of antibody Product replacements depend on product availability
Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052 Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice DevelopmentCustom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrichcomsafetycenter
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NYD77157-510122
1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
sigma-aldrichcom
Enabling Science to Improve the Quality of Life
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Order sigmacomorder Technical service sigmacomtechinfo sigmacomlifescience 23
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries SAGE and Where bio begins are trademarks of Sigma-Aldrich Co LLC
bioengineering
Biosynaptic
Advancements in neuroscience are yours to discover with targeted knockout rats from
SAGEtrade Labs Our comprehensive suite of rat models can help move your neuroscience research
forward Map your next breakthrough with smarter rat models for studying neurodegenerative
mood and affective disorders including seven new models of autism
Plot your pathway for breakthroughs in neuroscience with next-generation research models from SAGEtrade Labs
sageresearchmodelscom
OrderCustomer Service (800) 325-3010 bull Fax (800) 325-5052 Technical Service (800) 325-5832 bull sigma-aldrichcomtechservice DevelopmentCustom Manufacturing Inquiries (800) 244-1173 Safety-related Information sigma-aldrichcomsafetycenter
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NYD77157-510122
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copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
sigma-aldrichcom
Enabling Science to Improve the Quality of Life
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Sigma-Aldrichreg Worldwide Offices
NYD77157-510122
1121
copy2012 Sigma-Aldrich Co LLC All rights reserved SIGMA SAFC SIGMA-ALDRICH ALDRICH and SUPELCO are trademarks of Sigma-Aldrich Co LLC registered in the US and other countries FLUKA is a trademark of Sigma-Aldrich GmbH registered in the US and other countries PRESTIGE ANTIBODIES PANORAMA and FLAG are registered trademarks of Sigma-Aldrich Co LLC Where Bio Begins is a trademark of Sigma-Aldrich Co LLC Cy3 is a trademark of GE Healthcare Sigma brand products are sold through Sigma-Aldrich Inc Purchaser must determine the suitability of the product(s) for their particular use Additional terms and conditions may apply Please see product information on the Sigma-Aldrich website at wwwsigmaaldrichcom andor on the reverse side of the invoice or packing slip
World Headquarters 3050 Spruce St
St Louis MO 63103 (314) 771-5765
sigma-aldrichcom
Enabling Science to Improve the Quality of Life
ArgentinaFree Tel 0810 888 7446 Tel (+54) 11 4556 1472 Fax (+54) 11 4552 1698
AustraliaFree Tel 1800 800 097 Free Fax 1800 800 096 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500
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BelgiumTel (+32) 3 899 13 01 Fax (+32) 3 899 13 11
BrazilFree Tel 0800 701 7425 Tel (+55) 11 3732 3100 Fax (+55) 11 5522 9895
CanadaFree Tel 1800 565 1400 Free Fax 1800 265 3858 Tel (+1) 905 829 9500 Fax (+1) 905 829 9292
ChileTel (+56) 2 495 7395 Fax (+56) 2 495 7396
Peoplersquos Republic of ChinaFree Tel 800 819 3336 Tel (+86) 21 6141 5566 Fax (+86) 21 6141 5567
Czech RepublicTel (+420) 246 003 200 Fax (+420) 246 003 291
DenmarkTel (+45) 43 56 59 00 Fax (+45) 43 56 59 05
FinlandTel (+358) 9 350 9250 Fax (+358) 9 350 92555
FranceFree Tel 0800 211 408 Free Fax 0800 031 052 Tel (+33) 474 82 28 88 Fax (+33) 474 95 68 08
GermanyFree Tel 0800 51 55 000 Free Fax 0800 64 90 000 Tel (+49) 89 6513 0 Fax (+49) 89 6513 1169
HungaryTel (+36) 1 235 9055 Fax (+36) 1 235 9068
IndiaTelephone Bangalore (+91) 80 6621 9400 New Delhi (+91) 11 4358 8000 Mumbai (+91) 22 4087 2364 Pune (+91) 20 4146 4700 Hyderabad (+91) 40 3067 7450 Kolkata (+91) 33 4013 8000
Fax Bangalore (+91) 80 6621 9550 New Delhi (+91) 11 4358 8001 Mumbai (+91) 22 2579 7589 Pune (+91) 20 4146 4777 Hyderabad (+91) 40 3067 7451 Kolkata (+91) 33 4013 8016
IrelandFree Tel 1800 200 888 Free Fax 1800 600 222 Tel +353 (0) 402 20370 Fax + 353 (0) 402 20375
IsraelFree Tel 1 800 70 2222 Tel (+972) 8 948 4222 Fax (+972) 8 948 4200
Italy Free Tel 800 827 018 Tel (+39) 02 3341 7310 Fax (+39) 02 3801 0737
JapanTel (+81) 3 5796 7300 Fax (+81) 3 5796 7315
KoreaFree Tel (+82) 80 023 7111 Free Fax (+82) 80 023 8111 Tel (+82) 31 329 9000 Fax (+82) 31 329 9090
LuxembourgTel (+32) 3 899 1301 Fax (+32) 3 899 1311
MalaysiaTel (+60) 3 5635 3321 Fax (+60) 3 5635 4116
MexicoFree Tel 01 800 007 5300 Free Fax 01 800 712 9920 Tel (+52) 722 276 1600 Fax (+52) 722 276 1601
The Netherlands Tel (+31) 78 620 5411 Fax (+31) 78 620 5421New ZealandFree Tel 0800 936 666 Free Fax 0800 937 777 Tel (+61) 2 9841 0555 Fax (+61) 2 9841 0500
NorwayTel (+47) 23 17 60 00 Fax (+47) 23 17 60 10
PolandTel (+48) 61 829 01 00 Fax (+48) 61 829 01 20
PortugalFree Tel 800 202 180 Free Fax 800 202 178 Tel (+351) 21 924 2555 Fax (+351) 21 924 2610
RussiaTel (+7) 495 621 5828 Fax (+7) 495 621 6037
SingaporeTel (+65) 6779 1200 Fax (+65) 6779 1822
SlovakiaTel (+421) 255 571 562 Fax (+421) 255 571 564
South AfricaFree Tel 0800 1100 75 Free Fax 0800 1100 79 Tel (+27) 11 979 1188 Fax (+27) 11 979 1119
SpainFree Tel 900 101 376 Free Fax 900 102 028 Tel (+34) 91 661 99 77 Fax (+34) 91 661 96 42
SwedenTel (+46) 8 742 4200 Fax (+46) 8 742 4243
SwitzerlandFree Tel 0800 80 00 80 Free Fax 0800 80 00 81 Tel (+41) 81 755 2511 Fax (+41) 81 756 5449
ThailandTel (+66) 2 126 8141 Fax (+66) 2 126 8080
United KingdomFree Tel 0800 717 181 Free Fax 0800 378 785 Tel (+44) 1747 833 000 Fax (+44) 1747 833 313
United StatesToll-Free 800 325 3010 Toll-Free Fax 800 325 5052 Tel (+1) 314 771 5765 Fax (+1) 314 771 5757
VietnamTel (+84) 8 3516 2810 Fax (+84) 8 6258 4238
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