Re: Annexin V Positivity
Chris Reed (creed@pullman.com)
Tue, 04 Nov 1997 02:02:40 -0800
Robert Pyle wrote:
>
> I seem to remember a comment on the mail list regarding the induction of Annexin
> V positivity by manipulation of the cells of interest. Specifically, I remember
> the issue of freeze/thaw of banked cells leading to Annexin V positivity. Are
> there other cell manipulations that can lead to the same results? The reason I
> ask is that I am wondering if trypsinization of cells from culture plates will
> lead to Annexin V positivity like the freeze/thaw treatment? If so, it could
> answer some questions for us regarding some data we are fussing over.
>
> Any help or comments are enthusiastically welcomed.
>
> Haywood Pyle
> King Faisal Specialist Hospital and Research Center
> MBC #3, Box 3354, Riyadh 11211 Saudi Arabia
> e:pyle@kfshrc.edu.sa
> tel/fax: 966-1-464-7272 ext 32860
Haywood,
In my experience trypsinizing cells does *not* lead to Annexin binding
above background levels of 3% or so. I work with adherent cell lines and
have no problem telling the difference between cells treated with an
apoptosis inducer or an untreated negative control. I trypsinize for 7-8
minutes and then quench the reaction with media containing FBS or BCS
(1%).
Chris Reed
Senior in Genetics and Cell Biology
Washington State University
Pullman Washington