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>
>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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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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