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Updated: Jul 7, 2026
Hybrid Clear/Blue Native Electrophoresis for the Separation and Analysis of Mitochondrial Respiratory Chain Supercomplexes
Published on: May 19, 2019
Two-dimensional blue native/blue native polyacrylamide gel electrophoresis for the characterization of mitochondrial
Stephanie Sunderhaus1, Holger Eubel, Hans-Peter Braun
1Abt. Angewandte Genetik, Naturwissenschafiliche Fakultät, Universität Hannover, Germany.
Abstract:
Blue native polyacrylamide gel electrophoresis (BN-PAGE) employs the dye Coomassie for the labeling of proteins and protein complexes under native conditions. Electrophoresis under native conditions subsequently allows resolution of proteins and protein complexes according to their molecular mass. BN-PAGE can be combined with second gel dimensions. Best known is the two-dimensional (2D)-BN/sodium dodecyl sulfate (SDS)-PAGE system, which allows resolution of subunits of protein complexes. A 2D-BN/BN-PAGE system was developed that proved useful for investigating the substructure of protein complexes and protein supercomplexes. The basis of this 2D system is a variation of the conditions used for the two BN gel dimensions. Here, we present a basic protocol for the analysis of mitochondrial fractions by 2D-BN/BN-PAGE. Because both el dimensions are carried out under native conditions, the 2D-BN/BN system is compatible with in-gel enzyme activity staining.
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