An improved SDS-polyacrylamide gel electrophoresis for resolution of peptides in the range of 3.5-200kDa.

Abstract

Resolution of a wide range of polypeptides, 3.5-200kDa, on a single low acrylamide and cross linkage gel of 7.7% T, 2.6% C is described here. Laemmli's (4) original discontinuous SDS polyacrylamide gel electrophoresis (SDS-PAGE) system is modified by increasing the ionic strength of both stacking and resolving gels, and replacing the usual glycine buffer with a tricine cathode buffer as described by Schagger and von Jagow (7). This system offers the advantage of a wide range of protein fractionation, with sufficient band resolution, on a single, low acrylamide concentration and cross linkage gel. Moreover, increased gel ionic concentration allows higher protein and salt load, and renders this system suitable for preparative work.