Investigating Bak/Bax activating conformation change by immunoprecipitation.
Activation of both Bax and Bak during apoptosis involves significant conformation change. Investigation of this phenomenon by immunoprecipitation (IP) requires a detergent such as CHAPS that does not induce significant conformation change. IP with conformation-specific Bax or Bak antibodies is observed in CHAPS only following an apoptotic stimulus, whereas the same antibodies will immunoprecipitate from both nonapoptotic and apoptotic cells in the presence of Triton X-100. Thus, the latter detergent can serve as a positive control for IP, as described here.
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; D003902:Detergents; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods*; D017830:Octoxynol; 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; D003902:Detergents; D050998:bcl-2 Homologous Antagonist-Killer Protein; D051028:bcl-2-Associated X Protein; D017830:Octoxynol