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 quantitating proteins.
The complete V3 Western Workflow incorporates traditional blotting techniques with innovative tools, such as TGX Stain-Free™ Precast Gels, the Trans-Blot® Turbo™ Transfer System, and the ChemiDoc™ MP and ChemiDoc Imaging Systems, 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.
5 Steps
- Separate proteins — Mini-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.
- Visualize proteins — protein separation can be visualized and confirmed using stain-free technology, which covalently binds proteins, after a 1-minute activation on a ChemiDoc MP or ChemiDoc 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.
- 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.
- Verify protein transfer — the ChemiDoc MP or ChemiDoc Imagers, paired with stain-free technology, enable instant verification of protein transfer. After transfer, immunodetect your protein of interest using Precision Antibodies coupled with Clarity or Clarity Max ECL Substrates.
- Validate and quantitate — the fifth step, available only with a ChemiDoc MP or ChemiDoc Imager and Image Lab™ Software, may be used to validate western blotting data via total protein normalization. When normalizing to total protein, stripping and reprobing is no longer necessary. Image Lab Software gives quantitative results instead of relative normalization.
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