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We've recently started doing a lot of (flow) work with the oncor
Apoptag Kit; we've worked out conditions which give pretty good results
(thymocytes, human PBL, various cell lines), and the anti-digoxigenin FITC
labelling is adequate though I would like brighter staining for some
purposes. But my question is: the percentage of cells (apoptotic)
staining for dig-FITC is always significantly higher than percentages of
sub diploid "apoptotic" cells, based on P.I. analysis with ModFit (v1.01
for FACStation... aaaggghhh!) or other methods of analysis. I assume the
sub diploid DIG-FITC negative peak is necrosis, but what of DIG-FITC
positive sub diploids? Is PI just a grossly insensitive measure of
apoptosis (relative to the Apoptag kit) and how do you reconcile the two?
I was hoping someone would have a quick answer and or point me in
the direction of a few references.
T H A N K S !
CLAUDE CANTIN
cantinc@ircm.umontreal.ca
CYTOMETRIE EN FLUX / FLOW CYTOMETRY
INSTITUT DE RECHERCHES CLINIQUES DE MONTREAL
TEL. 514-987-5608
FAX 514-987-5633
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