Electrophoretic isolation of membrane proteins from acrylamide gels

Abstract

Protocols for the optimal resolution of membrane and watersoluble proteins in SDS-denatured state (Tricine SDS-PAGE and Blue Tricine SDS-PAGE; Laemmli SDS-PAGE and Blue Laemmli SDSPAGE) and in the native state (Blue Native PAGE) are presented. The protocols for protein recovery from these gels include techniques of electroelution and electroblotting optimized to the type of the preceding electrophoresis system. Native and denatured proteins thus are obtainable in near quantitative yield in soluble and in immobilized form. These techniques can optionally be performed in the milligram range, e.g., for the use of immunization and N-terminal protein sequencing, or in the analytical range.