Detection of apoptosis

From: DARZYNKIEWICZ ZBIGNIEW (DARZYNK@nymc.edu)
Date: Tue Apr 02 2002 - 15:36:46 EST


> 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
>
>
>



This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:05 EST