STLCC_CPLS;Morrison 3/28/2011Page 1
Prepared by: Bob Morrison
STLCC_CPLS, Instrumentation Specialist
Dec 08, rev Feb 09
BioRad IEF BioRad PowerPac HC
BioRad Criterion Cell
BioRad Protean II xi Proteomics
2D Gel Electrophoresis
Protocol
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Proteomics : BioRad 2-D
Systems Overview
Link to BioRad IEF Methods
and Products Manual (pdf)
BioRad PowerPac HC
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Proteomics : BioRad Protean IEF Cell, Basic Features
The PROTEAN IEF cell is a dedicated instrument for running IPG strips.
High-capacity running platform accommodates up to twenty-four 7 cm strips or twelve 11, 17, 18, or 24
cm strips to increase efficiency and throughput
Precision temperature control with a Peltier-cooled platform ensures reproducible focusing and an
expanded operating range of 10–25°C
Integrated 10,000 V, 2.4 mA power supply helps achieve the desired volt-hours faster when maximum
voltage is reached
Disposable rehydration/equilibration trays simplify the transfer of rehydrated strips into the focusing tray,
and have the added benefit of ensuring that residual sample buffer does not interfere with the current
path through the strips and adversely affect results
Tray options simplify handling. Each tray holds 1–12 IPG strips in individual numbered channels. For a
fully automated approach, rehydrate and run in the focusing tray. To increase throughput, rehydrate
strips in a rehydration/equilibration tray while running another set of IPG strips in the focusing tray in the
cell
Preset methods and real-time editing give maximum flexibility. The text-based user interface allows easy
programming of methods and provides a continuous update of run progress
Optional thermal printer records critical run data and provides a permanent record of the programmed
method and all run events. Run data are collected via the RS-232 serial port
Link to BioRad IEF Cell Instruction Manual (pdf)
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Proteomics : BioRad PowerPac HC Basic Features
PowerPac HC power supply, with 3.0 A of current and 300 W of power. In
addition to running standard horizontal agarose and vertical polyacrylamide
gels, the PowerPac HC power supply easily handles all your blotting
applications. To help you optimize every run, the PowerPac HC power
supply provides fully adjustable voltage, current, and power controls.
Runs 4 cells simultaneously
Timer-controlled
Fully programmable
CE and EN-61010 safety compliant
LCD screen shows all parameters during run
Case is stackable
Link to BioRad PowerPac Instruction Manual (pdf)
Proteomics: ReadyPrep 2-D Starter Kit Instructions
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Link to BioRad
ReadyPrep 2-D
Instruction Manual (pdf)
Proteomics: Protean II xi
Remove Comb & Clamps
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Remove Comb carefully from
one end
Remove sandwich clamp from
end while holding plate
Proteomics: ProteanII xi
Remove Clamps and Rinse
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Remove other sandwich clamp
Thoroughly rinse edge of wells
with DI water.
Proteomics: ProteanII xi
Remount clamps
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Remount plates in sandwich
clamps and lower assembly
into upper slots first
While pressing down and
forward lock sandwich clamps
into fixture
Proteomics: ProteanII xi
Load plates to Cell
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Install other plate on sandwich
clamps, ensure that the
indented edge will be facing in
toward the fixture center.
Load both plates with
sandwich clamps into cell
center fixture.
Proteomics: Protean II xi
Fill outer wells with Buffer
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Load the entire sandwich clamp
assembly into the cell
Fill outer wells with buffer to a
level of about one half full
(must cover lower plate edge)
Proteomics: Protean II xi
Fill inner well with Buffer
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Fill the inner well (between the
plates) completely full of buffer
Approximate Fill level for buffer
in outer wells
Proteomics: Mini-Protean Tetra Cell
Unlock Plates and remove
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Cell with plates and comb
Open “doors” to forward
position to release plates
Proteomics: Mini Protean Tetra Cell
– Remove comb
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Remove the comb carefully
from one end
Approximate Fill level for buffer
in outer wells
Proteomics: Mini Protean Tetra cell
Load Front side plate
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In-step side of plate with
channels goes toward inner
Wet seal line with buffer and
insert front plate.
Proteomics: Mini Protean Tetra cell
Complete Plate assembly
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Hold plates in position before
closing door clamps
Proteomics: Mini Protean Tetra cell
Load Plate Assembly in Cell
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Close door end clamps
Add plate assembly to cell
fixture
Proteomics: Mini Protean Tetra Cell
Fill wells with Buffer
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Fill outer portion of cell with
buffer to about 1/3rd level
Completely fill inner well area
with buffer
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Link to BioRad
2-D In a Day
Protocol (pdf)
Proteomics : BioRad 2D in a Day, Step 1
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Link to BioRad
2-D In a Day
Protocol (pdf)
Proteomics : BioRad 2D in a Day, Steps 2,3,4
Proteomics: ReadyPrep 2-D Flowchart
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Proteomics: ReadyPrep 2-D, Sample Prep, Rehydration
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Proteomics: ReadyPrep 2-D, Load Trays, Remove IPG Cover Strip
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Proteomics: ReadyPrep 2-D, IPG Strips onto Samples
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Proteomics: ReadyPrep 2-D, Isoelectric Focusing
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Proteomics: ReadyPrep 2-D, Wetting,Drain, Focus Tray
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Proteomics: ReadyPrep 2-D, Focus Tray Program
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Proteomics: ReadyPrep 2-D, 4.4 Completing IEF, Staining
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Proteomics: Trans-Blot Cell Assembly
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Proteomics: Trans-Blot- Prepare for Blotting, Steps 1-3
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Proteomics: Trans-Blot , Preparation, Steps 4-6
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Proteomics: Trans-Blot , Preparation, Steps 7-9
Proteomics: ReadyPrep2D, E.coli after stain, Prep for SDS-PAGE
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Proteomics: ReadyPrep 2-D, SDS-PAGE Buffer I and II, IPG
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Proteomics: ReadyPrep2D, SDS-PAGE
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Proteomics: ReadyPrep2D, IPG Strip to SDS-Page GEL
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Proteomics: ReadyPrep2D, Into IPG well with Melted Agarose
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Proteomics: ReadyPrep2D, 4.9 Staining
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Proteomics: ReadyPrep2D, Imaging on Densiometer
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