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Open Forum on Methodology

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How to do annexin V staining?

15 December 2000

Chuan-Wei Jang

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.

C.W. Jang

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Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882