Western Blot: Proteins Separating Technique, Protocol, Theory and Trouble Shooting

Abstract

Western blotting is used to visualize proteins that have been separated by gel electrophoresis. It is an important technique used in cell and molecular biology. Specific proteins can be identified from a complex mixture of proteins extracted from cells. The western blot includes three major steps: A. Separation by size B. Transfer to a solid support and C. Marking target protein using a proper primary and secondary antibody to visualize This technical article will attempt to explain the technique in detail to help researcher in getting good results and theory behind western blot, and offer some ways to troubleshoot.