Preparation of COS7 cells for FACS analysis

M. Johan Broekman, PhD mjbroek at
Mon Nov 24 21:14:10 EST 1997

We have no problems with lifting the cells with EDTA/BSA, especially if they
have only grown for 1-2 days.  Try putting the dish on wet ice, and light
scraping with a "rubber policeman".

Jun-Ichi Aikawa wrote:

> Hello. Does anyone try to prepare COS7 cells for analysis of cell-surface
> antigen with FACS? When I treated cells with PBS/5mM EDTA(pH 7.2) to
> prepare single-cell suspension, the efficiency is nearly 20-40% and most of
> cells still remains undispersed.  I also tried the increase of EDTA
> concentration (up to 25mM) and the addition of BSA(1%), but no improvement
> was observed. What buffer is suitable for this study? Any comments and
> suggestions will welcome.
> Jun-ichi Aikawa, Ph.D.
> University of California, San Diego

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