Detection of Bak/Bax activating conformation change by intracellular flow cytometry

G Dewson - Cold Spring Harbor Protocols, 2015 - cshprotocols.cshlp.org
G Dewson
Cold Spring Harbor Protocols, 2015cshprotocols.cshlp.org
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 …
Abstract
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.
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