Two-dimensional electrophoretic analysis of erythrocyte membranes.

摘要: In an effort to maximize the amount of genetic information that can be extracted from a blood sample, we investigated use two-dimensional polyacrylamide-gel electrophoresis (PAGE) resolve protein constituents erythrocyte membrane. Lyophilized membranes were dissolved in various concentrations urea, NP-40 detergent, and mercaptoethanol subjected PAGE by modification O'Farrell procedure, with ISO-DALT apparatus. More than 600 spots visible silver-stained gels under conditions excluded specific cytoskeleton components, including spectrin actin. The reproducibility pattern depended highly on precise composition solubilization mixture. Poor resolution was observed presence actin other proteins high molecular mass (spectrin bands 1 2) when used urea solubilized entire large number polypeptides could not attributed proteolysis, because addition proteolytic inhibitors membrane wash solutions did alter gel. also appear include cytosol because, except for globin, none five purified lysate gels. We conclude provides powerful tool study non-cytoskeletal proteins.