The Tris-Glycine-SDS Running Buffer is the most commonly used buffer for the separation/resolution of proteins in their denatured state during SDS-PAGE. It serves as both the anode and cathode buffer during gel electrophoresis and helps to regulate the pH of the electrophoresis system. When diluted to 1X, this buffer regulates pH levels at a pH of about 8.6, which provides reproducible separation of a wide range of proteins into well-resolved bands, depending upon the gel percentage used.