Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) Technology

Blue native polyacrylamide gel electrophoresis (BN-PAGE) is a separation method with a higher resolution than gel filtration or sucrose density ultracentrifugation that can be used to analyze abundant, stable multiprotein complexes (MPCs) from 10 kD to 10 MD. BN-PAGE uses gradient polyacrylamide gel for separation. During the process of separation, the native proteins and complexes tend to migrate according to the pore size of the gradient gel till they reach the pore size limit point. In contrast to immunoprecipitation and two-hybrid approaches, it allows the determination of the size, the relative abundance, and the subunit composition of an MPC. Applications of BN-PAGE include isolation and fractionation of membrane proteins, fractionation and identification of mitochondrial proteins, estimation of the native mass of the complex and its polymeric or oligomeric state, stoichiometry determination of a protein complex, 2D crystallization, electron microscopy, in-gel activity assays, native electroblotting, and immunodetection.