Proteomics 2DGel Electrophoresis

BioRad IEF
BioRad PowerPac HC
BioRad Protean II xi Proteomics 2D Gel Electrophoresis Protocol
BioRad Criterion Cell

Prepared by: Bob Morrison STLCC_CPLS, Instrumentation Specialist Dec 08, rev Feb 09
STLCC_CPLS;Morrison 3/28/2011
Page 1
Proteomics : BioRad 2-D Systems Overview

Link to BioRad IEF Methods and Products Manual (pdf)
BioRad PowerPac HC
Page 2
Proteomics : BioRad Protean IEF Cell, Basic Features

Link to BioRad IEF Cell Instruction Manual (pdf)
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 1025C 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 112 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
STLCC_CPLS;Morrison 3/28/2011 Page 3
Proteomics : BioRad PowerPac HC Basic Features

Link to BioRad PowerPac Instruction Manual (pdf)
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
Page 4
Proteomics: ReadyPrep 2-D Starter Kit Instructions
Link to BioRad ReadyPrep 2-D Instruction Manual (pdf)
Page 5
Proteomics: Protean II xi Remove Comb & Clamps

Remove Comb carefully from one end
Remove sandwich clamp from end while holding plate
Page 6
Proteomics: ProteanII xi Remove Clamps and Rinse
Remove other sandwich clamp
Thoroughly rinse edge of wells with DI water.
Page 7
Proteomics: ProteanII xi Remount clamps
Remount plates in sandwich clamps and lower assembly into upper slots first
While pressing down and forward lock sandwich clamps into fixture
Page 8
Proteomics: ProteanII xi Load plates to Cell
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.
Page 9
Proteomics: Protean II xi Fill outer wells with Buffer
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)
Page 10
Proteomics: Protean II xi Fill inner well with Buffer

Fill the inner well (between the plates) completely full of buffer
Approximate Fill level for buffer in outer wells
Page 11
Proteomics: Mini-Protean Tetra Cell Unlock Plates and remove
Cell with plates and comb
Open doors to forward position to release plates
Page 12
Proteomics: Mini Protean Tetra Cell Remove comb

Remove the comb carefully from one end
Approximate Fill level for buffer in outer wells
Page 13
Proteomics: Mini Protean Tetra cell Load Front side plate
In-step side of plate with channels goes toward inner
Wet seal line with buffer and insert front plate.
Page 14
Proteomics: Mini Protean Tetra cell Complete Plate assembly
Hold plates in position before closing door clamps
Page 15
Proteomics: Mini Protean Tetra cell Load Plate Assembly in Cell

Close door end clamps
Add plate assembly to cell fixture
Page 16
Proteomics: Mini Protean Tetra Cell Fill wells with Buffer

Fill outer portion of cell with buffer to about 1/3rd level
Completely fill inner well area with buffer
Page 17
Proteomics : BioRad 2D in a Day, Step 1
Link to BioRad 2-D In a Day Protocol (pdf)

STLCC_CPLS;Morrison 3/28/2011 Page 18
Proteomics : BioRad 2D in a Day, Steps 2,3,4
Link to BioRad 2-D In a Day Protocol (pdf)
Page 19
Proteomics: ReadyPrep 2-D Flowchart
Page 20
Proteomics: ReadyPrep 2-D, Sample Prep, Rehydration
Page 21
Proteomics: ReadyPrep 2-D, Load Trays, Remove IPG Cover Strip
Page 22
Proteomics: ReadyPrep 2-D, IPG Strips onto Samples
Page 23
Proteomics: ReadyPrep 2-D, Isoelectric Focusing
Page 24
Proteomics: ReadyPrep 2-D, Wetting,Drain, Focus Tray
Page 25
Proteomics: ReadyPrep 2-D, Focus Tray Program
Page 26
Proteomics: ReadyPrep 2-D, 4.4 Completing IEF, Staining
Page 27
Proteomics: Trans-Blot Cell Assembly
Page 28
Proteomics: Trans-Blot- Prepare for Blotting, Steps 1-3
Page 29
Proteomics: Trans-Blot , Preparation, Steps 4-6
Page 30
Proteomics: Trans-Blot , Preparation, Steps 7-9
Page 31
Proteomics: ReadyPrep2D, E.coli after stain, Prep for SDS-PAGE
Page 32
Proteomics: ReadyPrep 2-D, SDS-PAGE Buffer I and II, IPG
Page 33
Proteomics: ReadyPrep2D, SDS-PAGE
Page 34
Proteomics: ReadyPrep2D, IPG Strip to SDS-Page GEL
Page 35
Proteomics: ReadyPrep2D, Into IPG well with Melted Agarose
Page 36
Proteomics: ReadyPrep2D, 4.9 Staining
Page 37
Proteomics: ReadyPrep2D, Imaging on Densiometer
Page 38

Proteomics 2DGel Electrophoresis

Uploaded by

Document Information

Original Description:

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Proteomics 2DGel Electrophoresis

Uploaded by

Copyright:

Available Formats

BioRad IEF

BioRad Protean II xi Proteomics 2D Gel Electrophoresis Protocol

BioRad Criterion Cell

Proteomics : BioRad 2-D Systems Overview

Proteomics : BioRad Protean IEF Cell, Basic Features

Proteomics : BioRad PowerPac HC Basic Features

Proteomics: ReadyPrep 2-D Starter Kit Instructions

Link to BioRad ReadyPrep 2-D Instruction Manual (pdf)

Proteomics: Protean II xi Remove Comb & Clamps

Remove sandwich clamp from end while holding plate

Proteomics: ProteanII xi Remove Clamps and Rinse

Remove other sandwich clamp

Thoroughly rinse edge of wells with DI water.

Proteomics: ProteanII xi Remount clamps

Proteomics: ProteanII xi Load plates to Cell

Proteomics: Protean II xi Fill outer wells with Buffer

Load the entire sandwich clamp assembly into the cell

Proteomics: Protean II xi Fill inner well with Buffer

Approximate Fill level for buffer in outer wells

Proteomics: Mini-Protean Tetra Cell Unlock Plates and remove

Cell with plates and comb

Open doors to forward position to release plates

Proteomics: Mini Protean Tetra Cell Remove comb

Approximate Fill level for buffer in outer wells

Proteomics: Mini Protean Tetra cell Load Front side plate

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

Hold plates in position before closing door clamps

Proteomics: Mini Protean Tetra cell Load Plate Assembly in Cell

Add plate assembly to cell fixture

Proteomics: Mini Protean Tetra Cell Fill wells with Buffer

Completely fill inner well area with buffer

Proteomics : BioRad 2D in a Day, Step 1

Link to BioRad 2-D In a Day Protocol (pdf)

Proteomics : BioRad 2D in a Day, Steps 2,3,4

Link to BioRad 2-D In a Day Protocol (pdf)

Proteomics: ReadyPrep 2-D Flowchart

Proteomics: ReadyPrep 2-D, Sample Prep, Rehydration

Proteomics: ReadyPrep 2-D, Load Trays, Remove IPG Cover Strip

Proteomics: ReadyPrep 2-D, IPG Strips onto Samples

Proteomics: ReadyPrep 2-D, Isoelectric Focusing

Proteomics: ReadyPrep 2-D, Wetting,Drain, Focus Tray

Proteomics: ReadyPrep 2-D, Focus Tray Program

Proteomics: ReadyPrep 2-D, 4.4 Completing IEF, Staining

Proteomics: Trans-Blot Cell Assembly

Proteomics: Trans-Blot- Prepare for Blotting, Steps 1-3

Proteomics: Trans-Blot , Preparation, Steps 4-6

Proteomics: Trans-Blot , Preparation, Steps 7-9

Proteomics: ReadyPrep2D, E.coli after stain, Prep for SDS-PAGE

Proteomics: ReadyPrep 2-D, SDS-PAGE Buffer I and II, IPG

Proteomics: ReadyPrep2D, SDS-PAGE

Proteomics: ReadyPrep2D, IPG Strip to SDS-Page GEL

Proteomics: ReadyPrep2D, Into IPG well with Melted Agarose

Proteomics: ReadyPrep2D, 4.9 Staining

Proteomics: ReadyPrep2D, Imaging on Densiometer

You might also like