Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow™

Traditional western blot workflows require an appropriate sample load to detect both target proteins and loading control proteins. The discrepancy between these quantities often leads to oversaturated protein bands, forcing researchers to sometimes run two duplicate gels or titrate down the antibody concentration in order to get quantitative results.

The stain-free V3 Western Workflow requires knowledge of only the appropriate sample load for target protein detection and does not require reprobing of the blot for loading control proteins. More importantly, the stain-free blots exhibit greater linearity in the load range of 10–50 μg of cell lysate.

This application note describes an assay development step to determine the right load for the detection of target proteins before the actual western blot experiment is performed.