From: Alan Salkind (nepsy@fiona.umsmed.edu)
Date: Fri Apr 14 1995 - 13:56:42 EST
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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