POLY(ETHYLENE GLYCOL)METHACRYLATE ACRYLAMIDE COPOLYMER MEDIA FOR HYDROPHOBIC PROTEIN AND LOW-TEMPERATURE ELECTROPHORESIS

The properties of poly(ethylene glycol)methacrylate-acrylamide copolym er media (PEGMACs) were exploited in two ways: (i) in the first-dimens ional gel of two-dimensional electrophoresis (2-DE) of hydrophobic pro teins and (ii) for high speed, high-resolution electrophoresis at low temperatures. In the first application, improved resolution and yield of isoelectric focusing (IEF) separations for the hydrophobic protein zein was achieved compared to IEF in standard polyacrylamide gels. Thi s appears promising as a candidate approach for higher resolution 2-DE mapping of uncharacterized hydrophobic proteins. In the second applic ation, PEGMACs compatible with hydroorganic antifreeze buffer systems bellow cooling of the gels to low temperatures (-20 degrees C), which allowed greater current to be tolerated during electrophoresis. PEGMAC gels enabled us to perform sixfold faster electrophoretic separations and achieve threefold improved resolution of six standard proteins at the lower temperatures in a direct comparison with the normal sodium do decyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) system. If this approach is coupled with more precise instrumentation to cont rol low temperatures during electrophoresis, greater separation speeds and resolution may be anticipated.