Re: Annexin V/Propidium Iodide Staining for Adherent Cells

To circumvent this problem, determination of mitochondrial potential using
TMRE is a good choice and is followed by a number of laboratories. We have
used this to determine viability in  adherent epithelial cell lines
including insulinoma cell lines and MCF-7 (in preparation). The protocol
is simple and can be found in a number of publications including ours:
Jayaraman S, 2005; J Immunol Methods, 306:68-79.

Hope this helps.

SJ





On Thu, February 7, 2008 4:35 pm, Tavshanjian, Brandon wrote:
> Our lab has used a protocol for FITC-annexin V/PI staining to look at
> induction of
> apoptosis by flow which seems to work quite well in suspension BaF3 cells.
>  I'd like to
> use this technique in adherent cells (MCF7,MCF10A, other mammary
> epithelial cell lines),
> however, I've heard there are some issues with applying this technique to
> adherent cells
> (namely, that the cell detachment procedure may cause false-positive
> staining by annexin
> V).  I was wondering if anyone has a protocol like this that they've
> successfully used
> for adherent cells?
>
> ------------------------------------------------------
> Brandon Tavshanjian
> UCSF Chemistry and Chemical Biology
> Shokat Lab, Genentech Hall N-528
> (415) 514-4111
>
>
>
>
>
>
>
>


-- 
S. Jayaraman, Ph.D.
Associate Professor of Surgery
University of Illinois at Chicago
909 South Wolcott Avenue-704E MSB-M/C 790
Chicago, IL 60612
Phone: 312-355-5133
Fax: 312-355-1497