RE: Apoptosis detection

From: Hodge, Greg (HAEM) (hodgeg@mail.wch.sa.gov.au)
Date: Tue Apr 02 2002 - 18:20:33 EST


> ----------
> 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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