www.probes.com Reagents for Cell Biology and Imaging Molecular Probes Molecular Probes Technical Tips Nucleic Acid Stains Probes for the Cytoskeleton Mitochondrial Markers Probes for the Endoplasmic Reticulum and Golgi Apparatus Probes for Acidic Organelles and Endocytosis Probes for Long-Term Cell Tracing Lectin Conjugates Products for Special Applications Technical Tips Nucleic Acid Stains Probes for the Cytoskeleton Mitochondrial Markers Probes for the Endoplasmic Reticulum and Golgi Apparatus Probes for Acidic Organelles and Endocytosis Probes for Long-Term Cell Tracing Lectin Conjugates Products for Special Applications
Transcript
www.probes.com
Reagents for
Cell Biologyand Imaging
Molecular ProbesMolecular Probes
Technical Tips
Nucleic Acid Stains
Probes for the Cytoskeleton
Mitochondrial Markers
Probes for the Endoplasmic Reticulum and Golgi Apparatus
Probes for Acidic Organelles and Endocytosis
Probes for Long-Term Cell Tracing
Lectin Conjugates
Products for Special Applications
Technical Tips
Nucleic Acid Stains
Probes for the Cytoskeleton
Mitochondrial Markers
Probes for the Endoplasmic Reticulum and Golgi Apparatus
Probes for Acidic Organelles and Endocytosis
Probes for Long-Term Cell Tracing
Lectin Conjugates
Products for Special Applications
2
Product Color Coding
Fluorophore Color SelectionThroughout this brochure, we have assigned a color code to each fluorophore based on its peak
emission wavelength. We hope this will assist you in choosing the appropriate dye or combinationof dyes for your application. Figure 1 illustrates the spectral relationships between the color codeson a continuum from 400 to 700 nm.
Please note that the color code is only an approximation of what you would see if you looked atthe dye through the eyepiece of a standard fluorescence microscope; environmental factors and thefilters chosen to observe the dye affect the actual color observed through the eyepiece. Becausethe sensitivity of human vision falls off rapidly beyond ~650 nm, the fluorescence of our far-reddyes cannot easily be distinguished by eye. Consequently, the dyes in this range are arbitrarilyrepresented as magenta and purple.
How To Choose Dyes for Multicolor LabelingWhen you are using more than one fluorescent color, it is important to minimize the spectral
overlap (i.e., maximize the spectral separation) between probes. This will make it much easier toisolate their signals. If possible, select the brightest, most photostable probes for the lessabundant targets — green- and red-fluorescent dyes tend to be brighter than blue and far-red dyes.
To make an approximate assessment of the spectral separation between dyes, simply match thecolor codes of the products of interest to the spectrum in Figure 1. For a more accurate compar-ison, use the full emission spectra of the dyes (Figure 2). Spectra for most of our products can befound at our Web site, www.probes.com.
Figure 1.
400 500 600 700
Figure 2. A comparison of the spectral overlap between selected fluorophores. A) Normalized emission spec-tra for green-fluorescent Alexa Fluor 488 and red-fluorescent Alexa Fluor 594 dyes. Because these dyes areeasily distinguishable by eye and have minimal spectral overlap, they are a very suitable pair for multicolor ap-plications. B) Normalized emission spectra for Alexa Fluor 488 and orange-fluorescent Alexa Fluor 546 dyes.Although the peak emission colors of these dyes are easily distinguished by eye, the moderate spectral overlapindicates that there is still significant emission from Alexa Fluor 488 dye at the peak emission wavelength ofAlexa Fluor 546 dye. This signal “bleedthrough” can make separation of the dyes difficult in situations wherethe signal intensity of the Alexa Fluor 488 dye is much greater (due to an abundant target, for example) than thesignal intensity of the Alexa Fluor 546 dye. C) Normalized emission spectra for the orange-red–fluorescentAlexa Fluor 568 dye and Alexa Fluor 594 dye. Not only are the emissions from these dyes difficult to distin-guish by eye, but the spectral overlap is extensive, making this dye pair unsuitable for multicolor labeling underalmost all circumstances.
A B C
Molecular Probes 3
Antifade ReagentsMolecular Probes’ antifade reagents increase signal stability
and extend imaging time. Our antifade mounting media areavailable in these convenient kits:
• SlowFade Antifade Kit (S-2828) and SlowFade Antifade Kitwith DAPI (S-24635) — The SlowFade reagent is in a ready-to-use form and offers maximum bleaching protection in anonpermanent, glycerol-based mounting medium. Themounting medium is available either with or without DAPI,one of the most widely used nuclear counterstains.
Technical Tips• Selecting filters — The absorption and emission spectra of
fluorescent probes are generally asymmetrical. Most dyescontinue to exhibit significant absorption at wavelengths wellbelow their peak absorption and emit more light at wavelengthsthat are longer than their peak emission. In order to maximize thesignal from the fluorophore, it is usually best to choose filters thatare spectrally separated and slightly offset from the peakabsorption and emission wavelengths (Figure 3).
• Reducing photobleaching — Prolonged exposure to a light sourcewill eventually lead to photobleaching, an irreversible process thatdestroys the fluorophore. Because many imaging applicationsrequire extended periods of illumination, it is important to choosethe most photostable dye from among those that meet yourwavelength demands. Many of Molecular Probes’ proprietaryfluorophores, particularly our Alexa Fluor dyes, offer enhancedphotostability compared to similar fluorophores. An antifadereagent (see box below) can also be used to reducephotobleaching.
Figure 4. Bleaching profiles of A) fluorescein and B) Texas Red conjugates in cell samples. In thesephotobleaching experiments, human epithelium cells (Hep2) were probed with human anti-nuclear antibodiesand then developed for visualization with fluorophore-labeled secondary reagents. Identical samples weremounted in ProLong antifade reagent ( ), Product X (+) or medium containing no antifade reagent (o).Although these data were normalized, little or no quenching was observed for samples mounted with theProLong mounting medium.
Figure 3. Typical absorption (blue) and emission(green) spectra for a fluorescent dye, illustratingthe asymmetric nature of the curves. The shadedareas represent bandpass absorption and emis-sion filters optimized to maximize the signal fromthe dye.
• SlowFade Light Antifade Kit (S-7461) and SlowFade LightAntifade Kit with DAPI (S-24636) — Designed for applicationsin which signal intensity is critical, these alternate SlowFadeformulations offer photobleaching protection withoutcompromising initial signal brightness. The mounting mediumis available either with or without DAPI.
• ProLong Antifade Kit (P-7481) — This is our best all-aroundantifade preparation, offering outstanding photobleachingprotection in a permanent mounting agent (see Figure 4).
4
Nucleic Acid Stains
Cell-Permeant Nucleic Acid StainsMolecular Probes’ SYTO nucleic acid stains, which are available in several fluorescent colors,
readily penetrate most cell types. However, the SYTO dyes do not exclusively stain the nuclei of livecells — eukaryotic cells stained with SYTO dyes generally show both cytoplasmic and nuclearstaining. For researchers requiring a nuclear stain, Hoechst dyes are a better choice because theyare more DNA-selective.
SYTO Sampler KitsEven within a given color group of SYTO dyes,
there are structural differences between the dyes.These differences give rise to variations in thedyes’ excitation and emission maxima, and in theirrelative affinities for RNA and DNA. Researchershave discovered that, due to these variations,SYTO dyes have multiple applications, fromapoptosis markers 1 to RNA granule stains.2 Tohelp you decide which SYTO dye will work best foryour application, we have created the SYTONucleic Acid Stain Sampler Kits. These kitscontain color-grouped samples of our SYTO dyesto make it easier to find the best choice for aparticular cell or tissue type. Each of the SYTOdyes found in these kits is also available separately.
Hoechst 33342 and MitoTracker green FM
SYTO 16, MitoTracker Red and anti-tubulin antibody(detected with Alexa Fluor 350 dye-labeled secondary)
SYTO 13
TO-PRO-3 and Marina Blue streptavidin
Fluorescent Color Group Catalog Number
SYTO Blue Sampler Kit (SYTO dyes 40–45) S-11350
SYTO Green Sampler Kit #1 (SYTO dyes 11–16) S-7572
SYTO Green Sampler Kit #2 (SYTO dyes 20–25) S-7554
SYTO Orange Sampler Kit (SYTO dyes 80–85) S-11360
SYTO Red Sampler Kit (SYTO dyes 17, 59–64) S-11340
Abs* Em* Product Name Catalog Number
350 461 Hoechst 33342 H-1399, H-3570, H-21491
352 461 Hoechst 33258 H-1398, H-3569, H-21492
488 509 SYTO 13 S-7575
488 518 SYTO 16 S-7578
566 582 SYTO 85 S-11366
622 645 SYTO 59 S-11341
652 676 SYTO 62 S-11344
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
Molecular Probes 5
Nucleic Acid CounterstainsNucleic acid stains are especially valuable counterstains for
multicolor applications — an appropriate counterstain can lendcontrast to images and help define the spatial relationships betweencellular structures. Most of the counterstains listed in the tablebelow are impermeant to cells and, therefore, are generally used onfixed and permeabilized samples. Nucleic acid stains that are cell-impermeant may also be useful as dead-cell indicators because thedyes readily penetrate only those cells with compromised plasmamembranes.
We offer a broad selection of counterstains for immuno-histochemistry and other applications in which nucleic acids needto be visualized. These counterstains include:
• Standard counterstains — These dyes are commonly used fornuclear and chromosome staining. Due to its preference for DNAover RNA, DAPI exhibits very bright nuclear staining with little orno cytoplasmic background.
• TOTO and TO-PRO dyes — The TOTO and TO-PRO dyes all derivefrom the cyanine dye thiazole orange.3 TOTO dyes are dimers thathave a very high affinity for nucleic acids. They are suitable forsuch varied applications as counterstaining, electrophoresis, andsingle molecule imaging. TO-PRO dyes are monomeric counter-parts of the TOTO dyes. Although they have a somewhat loweraffinity for nucleic acids, the TO-PRO dyes still serve in many ofthe same applications as their dimeric cousins.
• SYTOX dyes — The SYTOX nucleic acid stains are chemicallysimilar to the SYTO dyes, but are cell impermeant. These dyesmay be used in multicolor applications or as dead-cell indicatorsin viability assays. SYTOX Green dye is, by far, the best green-fluorescent counterstain.
SYTOX Green Propidium iodide DAPI
Abs* Em* Product Catalog Number Applications and Notes
10. h tt p :/ / c o t t o n g e n o m e c e n t e r . u c d a v i s . e d u / p r o t o c o l s / p o p o 3 . h t m ; 11. Anal Chem 72, 4640 (2000); 12. Acta Histochem Cytochem 31, 297 (1998);
Phalloidin Conjugates for F-Actin StainingWe prepare numerous fluorescent derivatives of the cyclic peptide phalloidin for labeling the
cytoskeleton. Phalloidin derivatives selectively stain F-actin at nanomolar concentrations and arereadily water soluble, thus providing convenient labels for identifying actin in tissue sections andfixed-cell preparations.4 We have exploited the outstanding characteristics of our Alexa Fluor dyesto create ten different Alexa Fluor phalloidin conjugates that are superior in brightness andphotostability to all other spectrally similar phallotoxin conjugates.
Alexa Fluor 488 phalloidin
Oregon Green 488 paclitaxel
Polymerized tetramethylrhodamine–labeled tubulin
Abs* Em* Fluorophore Catalog Number
Alexa Fluor Phalloidin Conjugates
346 442 Alexa Fluor 350 A-22281
495 519 Alexa Fluor 488 A-12379
532 554 Alexa Fluor 532 A-22282
556 573 Alexa Fluor 546 A-22283
578 603 Alexa Fluor 568 A-12380
590 617 Alexa Fluor 594 A-12381
632 647 Alexa Fluor 633 A-22284
650 668 Alexa Fluor 647 A-22287
663 690 Alexa Fluor 660 A-22285
679 702 Alexa Fluor 680 A-22286
Other Phalloidin Conjugates
N/A N/A Biotin-XX B-7474
496 516 Fluorescein N-432
496 520 Oregon Green 488 O-7466
555 580 Rhodamine R-415
600 615 Texas Red-X T-7471
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
Molecular Probes 7
Antibodies for Immunolocalization of α-TubulinWhen used in conjunction with a secondary immunoreagent, Molecular Probes’ anti–bovine
α-tubulin monoclonal antibody enables researchers to visualize microtubules in fixed cells and infixed or frozen tissue sections from various species, including fruit flies,5 zebrafish 6 and humantumor cells.7 This mouse monoclonal antibody is available either unlabeled (A-11126) or as abiotin-XX conjugate (A-21371).
Probes for Studying the Cytoskeleton in Live CellsYou can visualize microfilament and microtubule dynamics in solution or in live cells using our
fluorescently labeled tubulin 8,9 and actin 10,11 monomers, which have been pre-tested and areready for microinjection.
The anti-cancer drug paclitaxel is a cell-permeant probe for labeling tubulin filaments in livecells. Unlike other fluorescent paclitaxel derivatives, the Oregon Green 488 paclitaxel conjugate ismade by derivatization of the 7β-hydroxy group of native paclitaxel, a strategy that permits theprobe to selectively bind to microtubules in live cells.12
Alexa Fluor 568
Alexa Fluor 546
Alexa Fluor 680
Alexa Fluor 633
Alexa Fluor 594
Reagents for Manipulating the CytoskeletonJasplakinolide (J-7473), a macrocyclic peptide isolated from the marine sponge Jaspis
johnstoni, is a potent inducer of actin polymerization in vitro 13 and competes with phalloidin foractin binding.14 Unlike other known actin stabilizers, such as phalloidins and virotoxins, jaspla-kinolide is somewhat cell-permeant and therefore may be useful for the manipulation of actinpolymerization in live cells. Latrunculin A (L-12370), isolated from a Red Sea sponge, is a toxinthat disrupts microfilament organization in cultured cells by binding to and sequesteringmonomeric G-actin.15
Alexa Fluor phalloidins with DAPI counterstain
Alexa Fluor 660Alexa Fluor 647
Abs* Em* Fluorophore Catalog Number
Actin (from rabbit muscle) Conjugates
495 519 Alexa Fluor 488 A-12373
578 603 Alexa Fluor 568 A-12374
N/A N/A Unlabeled A-12375
Tubulin (from bovine brain) Conjugates
511 530 Oregon Green 514 T-12391
555 580 Tetramethylrhodamine T-7460
N/A N/A Unlabeled T-7451
Paclitaxel Products
496 520 Oregon Green 488 P-22310
N/A N/A Unlabeled P-3456
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
8
Mitochondrial Markers
Mitochondrial Probes for Live CellsMolecular Probes’ wide selection of cell-permeant fluorescent mitochondrial stains includes
our proprietary MitoTracker and MitoFluor dyes. MitoTracker dyes contain a thiol-reactive moietythat allows them to be well retained after fixation. MitoFluor dyes serve as bright, photostablemitochondrial markers in live cells. Uptake of all these dyes depends somewhat upon the state ofthe mitochondrial membrane potential. However, MitoFluor Green and MitoTracker Green FMdyes produce bright, selective staining even in the absence of a potential difference across themitochondrial membrane.
Ratiometric Dyes for Mitochondrial Membrane PotentialJC-1 (T-3168) and JC-9 (D-22421) are cationic carbocyanine dyes that exist as green-
fluorescent monomers at low concentrations. Potential-driven mitochondrial accumulation ofthese dyes results in the formation of red-fluorescent “J-aggregates,” which exhibit a broadexcitation spectrum and a very narrow emission spectrum. Consequently, the ratioablefluorescence emission from these probes can be used as a sensitive measure of mitochondrialmembrane potential, particularly for the study of apoptosis.16-18
JC-9
MitoTracker Red
MitoFluor Red 594 and Hoechst 33342
MitoTracker Red, BODIPY FL phalloidin and DAPI
Abs* Em* Product Catalog Number Product Notes
484 519 DiOC6(3) D-273 Can be used to label both the endoplasmic reticulum 1
and mitochondria.2
489 517 MitoFluor Green M-7502 Stains mitochondria in both live and fixed cells.3
490 516 MitoTracker Green FM M-7514 Stains mitochondria in both live and fixed cells.3
507 529 Rhodamine 123 R-302 Useful for detecting membrane depolarization during
apoptosis.4
551 576 MitoTracker Orange M-7510 Stains mitochondria in live cells.5 Moderate retention
after fixation and permeabilization.
578 599 MitoTracker Red M-7512 Stains mitochondria in live cells. Good retention after
fixation and permeabilization.6
598 630 MitoFluor Red 594 M-22422 Optimal for use with 594 nm He-Ne laser sources.
680 710 MitoFluor Far Red 680 M-22423 Excitation bimodal with peaks at 650 and 690 in some
Probes for the Endoplasmic Reticulumand Golgi Apparatus
Cell-Permeant ER and Golgi ProbesThe endoplasmic reticulum (ER) and Golgi apparatus are primarily responsible for the proper
sorting of newly synthesized lipids and proteins in cells. In live cells, these structures can bestained with a variety of lipophilic probes and then distinguished based on their morphology. Toselectively stain the Golgi apparatus, you can use any of our fluorescent ceramide analogs, whichassociate preferentially with the trans-Golgi.
Probes for Immunodetection of the Golgi ApparatusWe offer two probes for the immunohistochemical and immunofluorescent detection of the
Golgi complex — anti–human golgin-97, mouse monoclonal antibody (A-21270) and lectin GS-IIfrom Griffonia simplicifolia. Golgin-97, a member of the granin family of proteins, is an integralmembrane protein localized on the cytoplasmic face of the Golgi complex.19,20 The GS-II lectinrecognizes nonreducing α- or β-linked N-acetyl-D-glucosamine residues 21 and is a highly selectivemarker for the Golgi complex.22 This lectin is available labeled with green-fluorescent Alexa Fluor488 dye (L-21415) or red-fluorescent Alexa Fluor 594 dye (L-21416), to provide the best andbrightest conjugates available.
ER-Tracker Blue-White DPXDiOC6(3), Alexa Fluor 633 phalloidin and DAPI
BODIPY FL C5-ceramide
Abs* Em* Product Catalog Number Product Notes
ER Probes
374 ~460 ER-Tracker Blue-White
DPX
E-12353 Very ER selective.1 Emission is highly environment
sensitive — UV longpass filters are recommended.
484 519 DiOC6(3) D-273 ER staining occurs after dye loss from mitochondria.2
Golgi Probes
466 536 NBD C6-ceramide N-1154
N-22651 †
Golgi selective in both live and fixed cells.3,4
503 510 BODIPY FL
C5-ceramide
D-3521
B-22650 †
Brighter alternative to NBD. Excimer formation occurs
in the trans-Golgi resulting in red-shifted emission.5
598 617 BODIPY TR ceramide D-7540 Useful for multicolor applications in both live and fixed
cells.6,7
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm. † In these products the ceramide is complexed to defatted BSA.
The defatted BSA facilitates cell labeling without requiring the use of organic solvents to dissolve the probe. 1. Mol Biol Cell 11, 1815 (2000);
Lysosomal ProbesOur LysoTracker probes are freely permeant to cell membranes and typically concentrate in
acidic organelles. The LysoTracker probes are available in several fluorescent colors, making themespecially suitable for multicolor applications. Furthermore, larger acidic compartments of cellsstained with LysoTracker Red DND-99 often retain their staining pattern following fixation withaldehydes. Researchers have used LysoTracker probes in a variety of applications, includingvisualization of acidic organelles in denervated skeletal muscle,23 assessment of acrosomalintegrity in cryopreserved bovine spermatozoa 24 and examination of lysosomal damage inTrypanosoma brucei after specific uptake of cytokine tumor necrosis factor-a.25
Fluorescent Bacteria and Yeast for Monitoring PhagocytosisFluorescent bacteria and yeast particles are proven tools for studying a variety of parameters
influencing phagocytosis, including the effects of anti-inflammatory drugs 26 and the role of theSrc family of tyrosine kinases in macrophages.27 Molecular Probes’ BioParticles products are aseries of fluorescently labeled, heat- or chemical-killed bacteria and yeast in a range of fluorescentcolors. We offer Escherichia coli (K–12 strain), Staphylococcus aureus (Wood strain withoutprotein A) and Saccharomyces cerevisiae (zymosan) BioParticles covalently labeled with a varietyof fluorophores, including two of our exceptional Alexa Fluor dyes.
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
Label Abs* Em* E. coli S. aureus S. cerevisiae
Fluorescein 494 518 E-2861 S-2851 Z-2841
Alexa Fluor 488 495 519 E-13231 S-23371 Z-23373
BODIPY FL 505 513 E-2864 S-2854 Z-2844
Tetramethylrhodamine 555 580 E-2862
Alexa Fluor 594 590 617 E-23370 S-23372 Z-23374
Texas Red 595 615 E-2863 Z-2843
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
Molecular Probes 11
Fluorescent Dextrans for Following EndocytosisThe molecular weights of our fluorescent dextran conjugates range from 3000 to
2,000,000 daltons. Many of them can be used to monitor the uptake and internal processingof exogenous materials by endocytosis.28,29 Our fluorescent dextrans may also be useful forstudies of endosome fusion 30 and vesicular morphology.31 Listed below is only a smallselection of some of our best dextran conjugates; please consult our Handbook of FluorescentProbes and Research Products for a complete product listing.
Membrane Probes for Following EndocytosisMolecular Probes’ membrane probes FM 1-43 (T-3163) and FM 4-64 (T-3166, T-13320) are
excellent reagents for monitoring endocytosis 32 and are particularly useful for investigating themechanisms of activity-dependent vesicle recycling in neurons from widely different species.33,34
These water-soluble dyes are virtually nonfluorescent in aqueous medium but become intenselyfluorescent when inserted into a membrane environment. In a neuron that is actively releasingneurotransmitters, these dyes become internalized within the recycled synaptic vesicles and thenerve terminals become brightly stained. The amount of FM 1-43 taken up per vesicle byendocytosis equals the amount of dye released upon exocytosis, indicating that the dye does nottransfer from internalized vesicles to endosome-like compartments during the recycling process.Both the absorption and fluorescence emission spectra of these dyes are significantly blue-shiftedin the membrane environment — FM 1-43 shows absorption and emission peaks at approximately480/565 nm 35 while FM 4-64 displays peaks near 515/640 nm.36
LysoTracker Yellow HCK-123 LysoTracker Red
MitoFluor 680, LysoTracker Green and Hoechst 33342
Label Abs* Em* 3000 MW 10,000 MW 70,000 MW
Cascade Blue 407 430 D-7132 D-1976
Fluorescein 494 518 D-3306 D-1820 D-1822
Alexa Fluor 488 495 519 D-22910
Tetramethylrhodamine 555 580 D-3308 D-1817 D-1818
Alexa Fluor 546 556 573 D-22911
Alexa Fluor 568 578 603 D-22912
Alexa Fluor 594 590 617 D-22913
Texas Red 595 615 D-3328 D-1863 D-1864
* Approximate absorption (Abs) and fluorescence emission (Em) maxima, in nm.
12
Probes for Long-Term Cell Tracing
Thiol-Reactive CellTracker DyesOur CellTracker dyes freely permeate the plasma membranes of live cells. Once inside the cell,
they bind to cellular components via the probes’ thiol-reactive chloromethyl group, trapping thetracers in the cytoplasm. Cell labeling with CellTracker dyes is stable for at least 24 hours afterloading, except in cells expressing ATP-dependent multidrug transporters, which rapidly excretedye-conjugated glutathione.37 The relatively long-term intracellular retention of CellTracker dyesmakes them ideal for any application requiring a persistent fluorescent label.
Amine-Reactive Cell TracersThere are only a few amine-reactive reagents that can passively diffuse into cells. These
compounds are colorless and nonfluorescent until their acetate groups are cleaved by intracellularesterases to yield the highly fluorescent, amine-reactive dye. Upon reaction with amine-containingresidues of intracellular proteins, these probes form dye–protein adducts that are well retained incells as they move and divide. Carboxyfluorescein diacetate succinimidyl ester (CFDA, SE —also referred to as CFSE in the literature) is the premier long-term tracer, with a signal that cansurvive for up to several weeks.38 The succinimidyl ester of SNARF-1 carboxylic acid acetate haspotential as a red-fluorescent alternative for tracing studies. However, the signal from SNARF-1 isless intense than that of carboxyfluorescein.
Lectin ConjugatesLectins bind to specific configurations of the various sugar molecules found in cellular
glycoproteins. Consequently, lectins are versatile primary detection reagents in histochemicalapplications.39 They are routinely used to identify specific cell and tissue types and to characterizedisease and injury states. Our Alexa Fluor lectin conjugates exhibit greater fluorescence intensityand photostability than any other similar dye conjugates.
Alexa Fluor 350 WGA, Alexa Fluor 568 phalloidin and SYTOX Green Alexa Fluor 488 Con A, Alexa Fluor594 WGA and Hoechst 33342 Alexa Fluor 488 GS-IB4 and
Fluorescent Nissl StainsNissl staining is a standard histological method for visualizing neurons in the brain and spinal
cord.40 Composed of ribosomal RNA associated with the rough endoplasmic reticulum in neuronalperikarya and dendrites, the Nissl substance redistributes within the cell body in injured or regen-erating neurons, providing a marker for the physiological state of the neuron. Our NeuroTracefluorescent Nissl stains are selective for the Nissl substance characteristic of neurons and providesignificantly more sensitivity than other histological Nissl dyes like toluidine blue or cresyl violet.
can be used to specifically identify neurons in human,41 mouse,42 avian,41 zebrafish,43 amphibian 44
and other tissues. The Hu antigen is an RNA-binding protein of the embryonic lethal abnormalvisual (ELAV) family, which includes the neuronal proteins HuD,45 HuC 40 46 and Hel-N1.47 Theantigen is expressed from the very early stages of neuronal development onward. We also offerthis antibody conjugated to biotin (A-21272).
Molecular Probes’ Alexa Fluor dyes are proprietaryand may be covered by U.S. Patent No. 5,696,157,U.S. Patent No. 6,130,101, or by one or more addi-tional pending U.S. or foreign patent applications.For research use only. Alexa Fluor, BioParticles,BODIPY, Cascade Blue, FM, LysoTracker, Marina Blue,MitoTracker, Oregon Green, ProLong, SlowFade,SNARF, SYTO, SYTOX, Texas Red, TO-PRO, TOTO,YO-PRO and YOYO are registered trademarks (®)of Molecular Probes, Inc. CellTracker, ER-Tracker,MitoFluor, NeuroTrace, and POPO are trademarks (™)of Molecular Probes, Inc.
Alexa Fluor 488 GS-IB4, NeuroTrace 530/615red-fluorescent Nissl stain and DAPI
NeuroTrace 530/615 red-fluorescent Nissl stain and DAPI
NeuroTrace 640/660 deep-red–fluorescent Nissl stain and DAPI
Abs* Em* Product Name Catalog Number
435 455 NeuroTrace 435/455 blue fluorescent Nissl stain N-21479
500 525 NeuroTrace 500/525 green fluorescent Nissl stain N-21480
Fluorescent ConjugatesMolecular Probes offers a wide variety of high-quality fluorescent bioconjugates that combine
our proprietary and conventional dyes with an assortment of biomolecules. Some of our newerconjugates, along with some interesting applications, are listed below.
Anti-Hu (detected with Alexa Fluor 546 dye–labeled secondary),Alexa Fluor 488 phalloidin and Hoechst 33342NeuroTrace 500/525 green-fluorescent
Visit Molecular Probesat Our Web Sitewww.probes.com
On the coverBovine pulmonary artery endothelial cells labeledwith MitoTracker Red CMXRos for mitochondria,BODIPY FL phallacidin for F-actin and DAPI fornuclei. This multiple-exposure image was acquiredusing DAPI, fluorescein and Texas Red bandpassoptical filter sets.
