Re: Apoptosis by FACS Protocols ?

Alan Salkind (nepsy@fiona.umsmed.edu)
Fri, 14 Apr 95 13:56:42 -0500

In response to Dr. Haury's inquiery, I've had good success using the Apotag
kit from Oncor. Very briefly, the fragmented 3'-OH DNA ends of intact cells
are labeled by digoxigenin-UTP (actually residues of digoxigenin-UTP are
catalytically added to the fragmented DNA) followed by incubation with
FITC-conjugated anti-digoxigenin. The cells can then be examined by flow
cytometry and/or fluorescence microscopy. I've found this method to be more
sensitive than DNA electrophoresis or propidium iodide for the detection of
apoptosis. Also, there is a commercially available Fas antibody available
from PanVera (Madison, Wisconsin), although I have no personal experience
with it.

Alan Salkind, M.D.
University of Mississippi Medical Center, Division of Infectious Diseases
Phone: 601-984-5560
Fax: 601-984-5565
e-mail: nepsy@fiona.umsmed.edu
>Hello,
>
>I'm looking for protocols and techniques to identify apoptotic cells on the
>FACS. Any methods or references are wellcome (I want to evaluate a bit
>what's already existing before deciding if I have to develop a new one...).
>
>Thanks a lot...
>
>Matthias
>
>------------------------
>Dr. Matthias Haury
>Flowcytometry Laboratory
>Department of Immunology
>Institut Pasteur Paris
>25 rue du Dr. Roux
>F-75724 PARIS France
>Tel: + 33 1 45 68 84 21
>Fax: + 33 1 45 68 86 39
>email: mhaury@pasteur.fr
>
>
>
>
ARS

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