Two-dimensional blue native/blue native polyacrylamide gel electrophoresis for the characterization of mitochondrial protein complexes and supercomplexes.

authored by
Stephanie Sunderhaus, Holger Eubel, Hans Peter Braun
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.

Organisation(s)
Institute of Plant Genetics
Type
Contribution to book/anthology
Pages
315-324
No. of pages
10
Publication date
2007
Publication status
Published
Peer reviewed
Yes
ASJC Scopus subject areas
Molecular Biology, Genetics
Electronic version(s)
https://doi.org/10.15488/11665 (Access: Open)
https://doi.org/10.1007/978-1-59745-365-3_23 (Access: Unknown)