Annexin controls

Tom Mc Closkey thomasm at nshs.edu
Thu Feb 20 13:02:33 EST 1997



--- On Tue, 18 Feb 1997 11:51:40 -0500  mamiller at 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 at nshs.edu 
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