--- On Tue, 18 Feb 1997 11:51:40 -0500 mamiller@wista.wistar.upenn.edu wrote: > >The way I see it, the BEST controls for an annexin experiment would >include a physiological negative control, a reagent/instrument negative >control, a positive control, and single-positive compensation controls >(PI only and annexin-FITC only) IF one is using seperate PI and annexin >solutions, as opposed to a pre-mixed kit. > While I agree with all the controls you listed, I would add one additional control [one which may not be presently available]. You need to assess how much of the annexin labelling is specific under the exact same culture conditions. i.e. the amount of nonspecific binding to the negative control [healthy cells] may not be representative of the the amount to the live cells in the conditions inducing apoptosis. With antibodies, an isotype control would be used [same fluor, same species, same isotype, etc.] So ideally you need an "annexin like molecule" or maybe annexin with the binding site deleted or something along those lines to answere this question. On your test sample, what do you compare it back to? -------------------------------------------------------- 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] 2/20/97 10:16:46 AM E-mail: thomasm@nshs.edu --------------------------------------------------------
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