Total Protein as a More Reliable and Reproducible Internal Control for Cancerous Liver Sample Western Blot Analysis
Dr. Mong-Lin Yang
Traditionally, protein normalization for Western Blot analysis has relied upon housekeeping proteins (HKPs) like β-actin and GAPDH. However, many studies have found HKPs to exhibit saturated signals and varied cellular expression levels that can lead to incorrect conclusions. Our work on comparing protein expressions between liver cancer samples versus normal liver samples encountered similar problems when employing β-actin and GAPDH as internal controls for protein normalization. In this project, we aim to test whether two commercially available total protein stains, Stain Free and No Stain, serve as better internal control for liver cancer protein analysis. Our result confirms total protein as the more reliable and reproducible internal control than HKPs. In addition, we identified No Stain as the preferred method between the two methods tested.