Bio-Rad's V3 Western Workflow is a five-step approach to streamlining your western blotting protocol. The complete V3 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, simplifying total protein quantitation analysis post-immunodetection to obtain results faster.
5 Step Workflow
1. 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.
2. Visualize proteins — Protein separation can be visualized and confirmed using stain-free technology, which covalently binds proteins, after 1 minute of activation on a 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 ChemiDoc MP imager paired with stain-free technology enables instant verification of protein transfer.
5. Validate and quantitate — The ChemiDoc MP imager and Image Lab™ software may be used to validate western blotting data via total protein normalization in conjunction with housekeeping proteins. By normalizing to total protein, stripping and reprobing is no longer necessary. Image Lab software gives quantitative results instead of relative normalization.