V3 Western Workflow™

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V3 Western Workflow - Western Blotting

Overview

V3 Western Workflow™
V3 Western Workflow™ with Stain-Free Gel Technology Tutorial

"Seeing is believing, measuring is knowing." Bio-Rad's V3 Western Workflow provides researchers with more confidence in the blotting process and in their results. This video demonstrates the fast, convenient, and transparent V3 Western Workflow process, how Image Lab™ software performs data normalization using total protein loading controls, and how stain-free gel technology is used to yield more reliable western blot results.

Ordering Information
  Mini-PROTEAN® TGX Stain-Free™ Precast Gels
 



 
Resolving
Gel %
8+1-Well
30 µl
10-Well
30 µl
10-Well
50 µl
12-Well
20 µl
15-Well
15 µl
IPG/Prep Well
7 cm IPG Strip
450 µl
 
7.5%
456-8029 456-8023 456-8024 456-8025 456-8026 456-8021
10%
456-8039 456-8033 456-8034 456-8035 456-8036 456-8031
12%
456-8049 456-8043 456-8044 456-8045 456-8046 456-8041
4–15% 456-8089 456-8083 456-8084 456-8085 456-8086 456-8081
4–20% 456-8099 456-8093 456-8094 456-8095 456-8096 456-8091
8–16% 456-8109 456-8103 456-8104 456-8105 456-8106 456-8101
Any kD™
456-8129 456-8123 456-8124 456-8125 456-8126 456-8121
 
Ordering Information
  Criterion™ TGX Stain-Free™ Gels
  12+2-Well 45 µl 18-Well 30 µl 26-Well 15 µl Prep+2-Well 800 µl IPG+1-Well 11 cm IPG Strip
 
Resolving Gel % 12+2-Well
45 µl
18-Well
30 µl
26-Well
15 µl
Prep+2-Well
800 µl
IPG+1-Well
11 cm IPG Strip
 
7.5% 567-8023 567-8024 567-8025
10% 567-8033 567-8034 567-8035
12% 567-8043 567-8044 567-8045
18% 567-8073 567-8074 567-8075 567-8072 567-8071
4–15% 567-8083 567-8084 567-8085 567-8082 567-8081
4–20% 567-8093 567-8094 567-8095 567-8092 567-8091
8–16% 567-8103 567-8104 567-8105 567-8102 567-8101
10–20% 567-8113 567-8114 567-8115 567-8112 567-8111
Any kD™ 567-8123 567-8124 567-8125 567-8122 567-8121
 

Bio-Rad's V3 Western Workflow is a five-step approach to streamlining your western blotting protocol. The V3 Western Workflow facilitates speed and validation at each step of a western blotting experiment — from running gels to quantifying proteins.

The complete V3 Western Workflow incorporates traditional blotting techniques with innovative tools, such as TGX Stain-Free™ precast gels, the Trans-Blot® Turbo™ system, and the ChemiDoc™ MP imager, to quickly check electrophoresis results and blot transfer quality prior to western blotting. This simplifies total protein quantitation analysis post-immunodetection and obtains results faster.

A simpler version of the V3 Western Workflow includes the Gel Doc™ EZ imager, a non-chemiluminescent capable system, in place of the ChemiDoc MP imager. This allows you to quickly check electrophoresis results and blot transfers while using traditional film detection for the chemiluminescent blot image.

5 Steps

  1. Separate proteinsMini-PROTEAN® TGX Stain-Free™ precast gels and Criterion™ TGX Stain-Free™ precast gels offer superior protein separation with fast run times and use standard Tris/glycine running buffers. TGX Stain-Free precast gels contain a unique technology formulated into the gel chemistry that enables rapid, high-quality separation of proteins in as little as 15 minutes.
  2. Visualize proteins — protein separation can be visualized and confirmed using stain-free technology, which covalently binds proteins, after a 1-minute activation on a Gel Doc EZ or ChemiDoc MP imager. This allows the immediate visualization of proteins across the whole gel without staining. Stain-free technology is a sensitive, time-saving alternative to traditional Coomassie staining, offering a streamlined workflow.
  3. Transfer proteins — the Trans-Blot Turbo system is a rapid protein transfer apparatus that can reduce transfer protocols to as little as 3 minutes while maintaining high-efficiency protein transfers across a wide range of molecular weights.
  4. Verify protein transfer — the Gel Doc EZ or ChemiDoc MP imagers paired with stain-free technology enable instant verification of protein transfer.
  5. Validate and quantitate — the fifth step, available only with a ChemiDoc MP imager and Image Lab™ software, may be used to validate western blotting data via total protein normalization. By normalizing to total protein, stripping and reprobing is no longer necessary. Image Lab software gives quantitative results instead of relative normalization.

Mini or Midi Gel Format

The complete V3 Western Workflow is offered in two gel formats with the ChemiDoc MP imager:

  • V3 Western Workflow complete system for mini precast gels (170-8292)
  • V3 Western Workflow complete system for midi precast gels (170-8293)

The modified V3 Western Workflow with the Gel Doc EZ imager is also offered in two gel formats:

  • V3 Western Workflow with Gel Doc EZ system for mini precast gels (170-8302)
  • V3 Western Workflow with Gel Doc EZ system for midi precast gels (170-8303)

More information

To select the best gel for your application, see the Mini Gel Selection Guide or the Midi Gel Selection Guide.

Visit the Applications and Technologies pages to learn more about the following:

V3 Western Workflow™ Complete System for Mini Gels

170-8292
ChemiDoc™ MP imager, Image Lab™ software, 50 Mini-PROTEAN® TGX Any kD Stain-Free™ precast gels, SDS-PAGE accessories, Mini-PROTEAN® Tetra cell, Trans-Blot® Turbo™ starter kit, 50 PVDF transfer packs for mini gels

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V3 Western Workflow™ Complete System for Midi Gels

170-8293
ChemiDoc™ MP imager, Image Lab™ software, 50 4–20% Criterion™ TGX Stain-Free™ precast gels, SDS-PAGE accessories, Criterion cell, Trans-Blot® Turbo™ starter kit, 50 PVDF transfer packs for midi gels

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V3 Western Workflow™ with Gel Doc™ EZ System for Mini Gels

170-8302
Gel Doc EZ imager with sample tray, Image Lab™ software, 20 4–20% Mini-PROTEAN® TGX Stain-Free™ precast gels, SDS-PAGE accessories, Mini-PROTEAN® Tetra cell, Trans-Blot® Turbo™ starter kit, 20 PVDF transfer packs for mini gels

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V3 Western Workflow™ with Gel Doc™ EZ System for Midi Gels

170-8303
Gel Doc EZ imager with sample tray, Image Lab™ software, 20 4–20% Criterion™ TGX Stain-Free™ precast gels, SDS-PAGE accessories, Criterion cell, Trans-Blot® Turbo™ starter kit, 20 PVDF transfer packs for midi gels

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Number Description Options
6216 Detection Buffer Formulations Protocol, Rev A Click to download
6218 Blot Stripping and Reprobing Protocol, Rev A Click to download
6427 Bio-Rad V3 Western Workflow, Rev A Click to download [ Add to Cart (Free) ]
6434 Western Blot Normalization Using Image Lab™ Software Quick Start Guide Click to download
6253 V3 Western Workflow Brochure with ChemiDoc MP Imager, Rev B Click to download [ Add to Cart (Free) ]
6088 V3 Western Workflow Brochure with Gel Doc EZ Imager, Rev B Click to download [ Add to Cart (Free) ]
6264 V3 Western Workflow Protocol Customer Sample, Rev B Click to download [ Add to Cart (Free) ]
RP0051 Stain-Free Approach for Western Blotting Click to download
2895 Protein Blotting Guide, Interactive PDF, Rev B Click to download
6133 ChemiDoc MP Brochure, Rev C Click to download [ Add to Cart (Free) ]
6039 Trans-Blot Turbo Transfer System Brochure, Rev D Click to download [ Add to Cart (Free) ]
6166 Multiplex Fluorescent Blot Detection: A Troubleshooting Guide, Rev A Click to download [ Add to Cart (Free) ]
6349 Total Protein Normalization Publications, Rev A Click to download
6350 Housekeeping Protein Normalization Publications, Rev A Click to download
6351 Stain-Free Technology Publications, Rev C Click to download
6359 Avoiding Housekeeping Protein Detection Saturation Protocol, Rev A Click to download
6361 Determining the Appropriate Film Exposure Time Protocol, Rev A Click to download
6362 Determining the Appropriate Sample Load for Western Blots Protocol, Rev A Click to download
6363 Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow Protocol, Rev A Click to download
6366 Validating the Expression Consistency of a Housekeeping Protein Protocol, Rev A Click to download
6376 General Protocol for Western Blotting, Rev A Click to download
6356 Traditional vs. V3 Western Workflow Click to download
6360 A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation, Rev B Click to download
6390 General V3 Western Workflow Blotting Protocol Click to download
RP0058 A Defined Methodology for Reliable Quantification of Western Blot Data Click to download
5781 Evaluation of the Criterion Stain Free Gel Imaging System for Use in Western Blotting Applications, Rev A Click to download [ Add to Cart (Free) ]