> ---------- > From: Zucker.Robert@EPAMAIL.EPA.GOV > Sent: Tuesday, 2 April 2002 3:44 > To: Cytometry Mailing List > Subject: Apoptosis detection > > Bob, > > I recently noticed Genzyme have a MoAb to ssDNA which they claim is > specific for Apoptosis. The antibody is designed for use with formamide, > which selectively denatures DNA in apoptotic cells. They induced necrosis > by hypothemia or detergent and DNA breaks by H2O2. These necrotic cells > were negative using this new MoAb but positive by TUNEL assay. They also > have an ELISA which they claim is sensitive to 500 apoptotic cells per > well. I have ordered the MoAb but have not tested its specificity. This is > possibly the only single technique that I have heard of to define > apoptotic cells by flow. Anyone else tried this technique ? > > Greg Hodge PhD > Senior Medical Scientist > Haematology Department > Women's and Children's Hospital > North Adelaide > South Australia 5006 > ph 08 81616089 > fax 08 81616043 > email hodgeg@mail.wch.sa.gov.au > -------------------------------------------------------------------------- > ------------------ > > > On 2nd April Robert Zucker wrote- > > > A colleague asked a simple question regarding a continous lymphoblast > system exposed to toxicants. What is the best assay or kit to detect > apoptosis? > After a short discussion on flow assays that included --permeability > endpoints, annexin, tunnel, and subg1, he decided on using the gold > standard of laddering. He wants a definitive test. I thought there > were more sensitive and more accurate flow cytometic assays to detect > apoptosis. I also told him he usually needs two tests to make a > conclusion reegarding apoptosis. Any opinions on what is the best and > easiest approach to detect apoptosis in a suspension cell culture? > > > > Bob > > Robert M. Zucker, PhD > U.S. Environmental Protection Agency > Office of Research and Development > National Health and Environmental Effects Research Laboratory > Reproductive Toxicology Division, MD 72 > Research Triangle Park, North Carolina, 27711 > Tel: 919-541-1585; fax 919-541-4017 > e-mail: zucker.robert@epa.gov > >
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