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 the molecular biology laboratories. However, western blot standard protocols may be unable to detect certain proteins depending on their size. When the protein of interest is out of 10-250 KDa range, its migration through the gel or transfer to the membrane are compromised, thus 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 (SVIP) and Max-Gene Associated Protein (MGA), whose molecular weight are 8 and 350 kDa, respectively. We expect that these adaptations may help researchers to improve their experiments in a cost-effective manner.