Protein analysis is an integral component of many research projects; understanding a protein of interest’s expression level or modification status can offer key mechanistic insights into the pathways that are active within a cell. To separate and analyze proteins, researchers most commonly use polyacrylamide gel electrophoresis (PAGE). While generations of scientists have used and improved protein gel electrophoresis, obtaining a clean protein gel with crisp bands remains a challenge.
To separate proteins by PAGE, scientists first choose between purchasing pre-cast gels or casting their own. While precast gels are convenient and show higher reproducibility rates than handmade gels, the latter gives researchers more flexibility to adjust a gel’s polyacrylamide concentrations to achieve optimal protein separation. Next, scientists must decide on a gel chemistry and buffer system for gel electrophoresis. Many researchers choose Tris-Glycine chemistry to separate proteins. In this method, scientists use Tris-HCl to create the protein gel while they ...
