Isolation of membrane protein complexes by blue native electrophoresis.

Abstract

Blue native PAGE is a discontinuous electrophoretic system that allows the separation of membrane protein complexes in a native, enzymatically active state with high resolution. Membrane protein complexes are solubilized by neutral, nondenaturing detergents like n-dodecyl-beta-D-maltoside. After addition of Coomassie G250 that binds to the surface of the proteins, separation of the negatively charged complexes according to molecular mass is possible. After electrophoresis the structure and function of the isolated protein complexes can be investigated.