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Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Article DOI: 10.3791/2164-v • 12:04 min • February 24th, 2011
February 24th, 2011 •
1,2,3, *2,3, *3
1Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, 2Centre for Biological Signalling Studies (bioss) and Biology III, Faculty of Biology, University of Freiburg, 3Department of Molecular Immunology, Max-Planck-Institute of Immunology and Epigenetics
* These authors contributed equally

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In this video, we describe the characterization of multiprotein complexes (MPCs) by blue native polyacrylamide gel electrophoresis (BN-PAGE). In a first dimension, dialyzed cellular lysates are separated by BN-PAGE to identify individual MPCs. In a second dimension SDS-PAGE, MPCs of interest are further subdivided to analyze their constituents by immunoblotting.

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Blue Native Polyacrylamide Gel Electrophoresis BN-PAGE Multiprotein Complexes Cell Signalling Protein-protein Interaction Networks Native Conditions Resolution Gel Filtration Sucrose Density Ultracentrifugation MPC Size Composition Relative Abundance Hydrodynamic Size And Shape Polyacrylamide Matrix Cellular Lysates Lysate Dialysis First Dimension BN-PAGE Second Dimension SDS-PAGE
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