Blue native PAGE

authored by

Ilka Wittig, Hans Peter Braun, Hermann Schägger

Abstract

Blue native PAGE (BN-PAGE) can be used for one-step isolation of protein complexes from biological membranes and total cell and tissue homogenates. It can also be used to determine native protein masses and oligomeric states and to identify physiological protein-protein interactions. Native complexes are recovered from gels by electroelution or diffusion and are used for 2D crystallization and electron microscopy or analyzed by in-gel activity assays or by native electroblotting and immunodetection. In this protocol, we describe methodology to perform BN-PAGE followed by (i) native extraction or native electroblotting of separated proteins, or (ii) a second dimension of tricine-SDS-PAGE or modified BN-PAGE, or (iii) a second dimension of isoelectric focusing (IEF) followed by a third dimension of tricine-SDS-PAGE for the separation of subunits of complexes. These protocols for 2D and 3D PAGE can be completed in 2 and 3 days.

Organisation(s)

Institute of Plant Genetics

External Organisation(s)

Goethe University Frankfurt

Type

Article

Journal

Nature protocols

Volume

1

Pages

418-428

No. of pages

11

ISSN

1754-2189

Publication date

27.06.2006

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)

Electronic version(s)

https://doi.org/10.1038/nprot.2006.62 (Access: Unknown)