Open Forum on Methodology
How to do annexin V staining?
15 December 2000
For detecting apoptosis, I tried annexin V staining. But after modifying reagent concentrations, temperature, and incubation time, I could not get the expected result! My experimental cell lines include Hepatoma 3B, 293T, MCF10A etc, but none seems to work. When I carried out DNA staining and FACS analysis, I can see many sub-G1 cells, but with the same sample, I see no annexin V-positive signal or get irrelevant results. I followed the usual method, harvested cells with trypsin, then wash twice, and stained. I also have asked others who used this method and they told that is a tricky!! Is it true? Are there key problems to be avoided? Thanks for your help.
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