--- On Fri, 23 Apr 1999 09:45:35 PDT Rita Cavaleiro <ritacavaleiro@hotmail.com> wrote: > I am not sure if >this high level represents unspecific labelling because there is no >wash after the adition of annexin. None of the protocols use this >wash. Could anyone tell me if there is any problem in doing this wash? We routinely wash with binding buffer after the annexin labeling step. > In addition, I am not sure about what control I have to use to put >the cut off. What is the best positive control to induce apotposis in >human PBMCs and in what concentrations ?. Crosslinking through CD4 with a GAM secondary or activation through CD3 will induce some level of cell death. A useful positive control is Jurkat cells treated with anti CD95 for a few hours. Good luck, Tom -------------------------------------------------------- Thomas W. Mc Closkey, Ph. D. Director, Flow Cytometry North Shore University Hospital Biomedical Research Center 350 Community Drive Manhasset, Long Island, New York 11030 ph: 516-562-4844 [office]; 516-562-1135/4641 [lab] fax: 516-562-2866 4/28/99 2:52:42 PM E-mail: thomasm@nshs.edu --------------------------------------------------------
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