Detection of Bak/Bax activating conformation change by intracellular flow cytometry.
Like the commonly used immunoprecipitation (IP) approach, this procedure for the detection of activated Bak or Bax by intracellular flow cytometry is based on the principle that Bak and Bax, during activation, expose occluded amino-terminal epitopes that can be recognized by conformation-specific antibodies. Flow cytometric analysis requires fewer cells and is less time-consuming than IP. Further, in contrast to IP, flow cytometry produces a quantifiable assessment of the percentage of cells containing activated Bak or Bax, which can be correlated with cell death.
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.
Grant
G
Dewson
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D011487:Protein Conformation; D050998:bcl-2 Homologous Antagonist-Killer Protein / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D051028:bcl-2-Associated X Protein / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein