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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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web