I have a client who has used this in virally infected cells with some success. However,there is not very good separation between negative and positive. I was told by a Pharmingen person that this was a problem with this particular lot of antibody. Also, it does have a lot of non-specific background over non-stained cells. Hopefully they will be coming out with the monoclonal version soon. Candace Enockson Medical University of South Carolina --On Tue, Sep 7, 1999 9:35 AM -0400 Maryalice Stetler-Stevenson <stetler@box-s.nih.gov> wrote: > > We also have tried this antibody in parallel with Annexin V without > success but haven't done a time course, etc to determine if there are > biological reasons this isn't working for us. I would be interested in > others experience with this antibody and any tricks people may have used > to get it to work better. > > Maryalice > >> Dear all >> >> We have been using a anti-Caspase-3 rabbit polyclonal ab conjugated >> with PE (this is supposed to bind to the active form only) by flow. >> We've looked for caspase-3 activation in camptothecin or etoposide >> treated Jurkatt cells (cytospins confirm apoptosis in these >> cells) after fixation and perm, without success. This ab is from >> Pharmingen. >> >> Does anyone have experience with looking for caspase-3 activation by >> flow ? Are there other antibodies available ? >> >> Any comments would be much appreciated. >> >> Fran Gibson >> Dept Haematology >> St. George's Hospital Medical School >> London > > Maryalice Stetler-Stevenson > Director Flow Cytometry Unit > Laboratory of Pathology, NCI, NIH
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