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

130.8K Views

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12:03 min

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February 24th, 2011

DOI :

10.3791/2164-v

February 24th, 2011

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Gina J. Fiala¹^,²^,³, Wolfgang W. A. Schamel*²^,³, Britta Blumenthal*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

Summary

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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

Chapters in this video

0:05

Title

1:21

Introduction

1:58

Preparation of Dialyzed Cell Lysate

4:48

Pouring of Blue Native-gels

7:04

Separation of Dialyzed Cell Lysate by Blue Native-PAGE

7:53

Second Dimension SDS-PAGE

9:25

Representative Results for Blue Native-PAGE Analysis of Multiprotein Complexes from Cellular Lysates

11:18

Conclusion

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