> Bob Zucker inquires about a definitive test of detection of apoptosis in > toxicology testing. > > My advice is to try the STATHMO-APOPTOSIS assay (Smolewski et al., > Cytometry, 47, 143, 2002). The assay has several advantages:. > (a) It is based on detection of caspases activation - activation of > caspases is considered to be a hallmark of apoptosis. (b) It is the only assay that measures cumulative apoptotic index (CAI). The CAI may be measured over long period of time - at least for 3 days. With any other assay apoptotic cells disintegrate and vanish from the analysis. Because in most situations the cells enter apoptosis asynchronously, the "snapshot" apoptotic index at a given, time point, as it is done with other methods, is inaccurate because it fails to detect the apoptotic cells that either already disintegrated or yet did not enter apoptosis. > (c) the assay is simple. Just add the labeled inhibitor (FLICA) into the > culture, then when harvesting cells, rince them twice and measure cell > fluorescence. It can be combined with cell fixation and bivariate analysis > of DNA content, DNA strand breakes etc. > Zbigniew Darzynkiewicz, M.D., Ph.D. Director, Brander Cancer Research Institute New York Medical College 19 Bradhurst Ave. Hawthorne, NY 10532 tel: 914-347-2801 fax: 914-347-2804 http://www.geocities.com/z_darzynkiewicz > > >
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