Unit

UNIT 6.10 Two-Dimensional Blue Native Polyacrylamide Gel Electrophoresis

  1. Wolfgang W.A. Schamel

Published Online: 1 MAR 2008

DOI: 10.1002/0471143030.cb0610s38

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Schamel, W. W. 2008. Two-Dimensional Blue Native Polyacrylamide Gel Electrophoresis. Current Protocols in Cell Biology. 38:6.10:6.10.1–6.10.21.

Author Information

  1. Max Planck-Institut fµr Immunbiologie und Universität Freiburg, Biologie III, Freiburg, Germany

Publication History

  1. Published Online: 1 MAR 2008
  2. Published Print: MAR 2008

Abstract

Multiprotein complexes play crucial roles in nearly all cell biological processes. Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) is a powerful method to study these complexes. It is a native protein separation method that relies on the dye Coomassie blue to confer negative charge for separation. It has a higher resolution than gel filtration or sucrose density ultracentrifugation and can be used for protein complexes from 10 kDa to 10 MDa. If a second-dimension SDS-PAGE is applied (two-dimensional BN/SDS-PAGE), the size, subunit composition, and relative abundance of the different multiprotein complexes can be studied. In recent years, there has been a large increase in the number of publications where BN-PAGE was used to study protein-protein interactions. Here, we give detailed protocols for the separation of multiprotein complexes by two-dimensional BN/SDS-PAGE and for a related technique to determine the stoichiometry of these complexes. Curr. Protoc. Cell Biol. 38:6.10.1-6.10.21. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • multiprotein complex;
  • native;
  • gel electrophoresis;
  • two-dimensional;
  • Coomassie blue;
  • protein-protein interaction