Bio-Rad’s TGX Stain-Free Gels use total protein normalization to produce a much greater linear dynamic range for measuring target protein levels (Figure 1). Housekeeping proteins such as β-actin, β-tubulin, or GAPDH are often very abundant in biological samples, which results in their signal being oversaturated compared to target proteins. Normalizing results to a total protein measurement corrects this problem, allowing a meaningful comparison even with low-abundance targets, and leads to far greater quantitative accuracy in measuring the levels of the protein of interest (Figure 2).
Bio-Rad’s V3 Western Workflow™ incorporates stain-free gel chemistry to streamline the western blotting protocol. This improved workflow, summarized below, saves time and increases accuracy and reliability throughout the western blotting process.
- Separate proteins using TGX Stain-Free Gels in as little as 15 min.
- Visualize protein separation in 1 min using the ChemiDoc™ Touch Imaging System.
- Transfer proteins efficiently and uniformly in 3 min using the Trans-Blot® Turbo™ Transfer System.
- Verify transfer efficiency quickly using the ChemiDoc Touch Imaging System.
- Validate western blot data by normalization and analysis using the stain-free blot image as the total protein loading control.