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
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