Blue Native PAGE and Antibody Gel Shift to Assess Bak and Bax Conformation Change and Oligomerization
- Cell Signalling and Cell Death Division, Walter and Eliza Hall Institute of Medical Research, Parkville, Melbourne, Victoria 3052, Australia;
- Department of Medical Biology, The University of Melbourne, Melbourne, Victoria 3010, Australia
Abstract
Blue native PAGE (BN-PAGE) uses Coomassie dye rather than denaturing SDS to provide a negative charge to proteins for electrophoresis. As such, it is a useful assay for investigating native supramolecular membrane complexes without the need for cross-linking. As Bak and Bax oligomers form in the mitochondrial outer membrane, and they can be efficiently monitored by BN-PAGE. Furthermore, BN-PAGE performed in conjunction with gel-shift using conformation-specific antibodies can provide additional information regarding the activation state of Bak or Bax in specific membrane complexes.
Footnotes
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↵1 Correspondence: dewson{at}wehi.edu.au
- © 2015 Cold Spring Harbor Laboratory Press
