How the West(ern) was won: Solutions for immunoblotting large and small proteins

Abstract

Relative protein quantification is a well-established technique in the vast majority of molecular biology laboratories. However, western blot standard protocols may not detect proteins of certain sizes. When the molecular weight of protein of interest is out of the 10–250 kDa range, its migration through the gel or transfer to the membrane is compromised, making its detection difficult. Here, we present a set of modifications of the standard working procedure for western blotting based on the experience working with small VCP-interacting protein and MAX-gene-associated protein, whose molecular weights are 8 and 350 kDa, respectively. We expect that these adaptations may help researchers to improve their experiments in a cost-effective manner.