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

verfasst von

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.

Organisationseinheit(en)

Institut für Pflanzengenetik

Typ

Beitrag in Buch/Sammelwerk

Seiten

315-324

Anzahl der Seiten

10

Publikationsdatum

2007

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Molekularbiologie, Genetik

Elektronische Version(en)

https://doi.org/10.15488/11665 (Zugang: Offen)
https://doi.org/10.1007/978-1-59745-365-3_23 (Zugang: Unbekannt)