UNIT 6.10 Two-Dimensional Blue Native Polyacrylamide Gel Electrophoresis
Published Online: 1 MAR 2008
Copyright © 2008 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAR 2008
- Published Print: MAR 2008
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.
- multiprotein complex;
- gel electrophoresis;
- Coomassie blue;
- protein-protein interaction