Re: Annexin V/PI and Adherent Cells

From: David L. Haviland (dhavilan@imm2.imm.uth.tmc.edu)
Date: Mon May 08 2000 - 15:38:21 EST


"Beverly E. Barton" wrote:
>
>    To the Cytometry List:
>
> I have successfully used GIBCO's Enzyme-Free Cell Dissociation Buffer
> for many adherent cell types with the annexin V/PI assay for apoptosis.
> The one cell type I was unsuccessful with is HUVEC.  For these cells,
> since I have to trypsinize the cells for the maximum viability, I use
> Molecular Probe's JC-1 mitochondrial stain to measure apoptosis by
> mitochondrial membrane depolarization.  This has worked very nicely.

Beverly:

What do you know of its composition?  Is it somewhat like the "Versene"
solution that is basicall PBS w/o Ca2+/Mg2+ but may have anywhere
between 0.5-5 mM EDTA?

The latter solution works well in my hands @ 5mM EDTA but as one might
guess - it doesn't do well for those cells that are moderately to very
well attached.

Many thanks,
David
===========
David L. Haviland, Ph.D., Asst. Prof. Immunology
University of Texas - Houston, H.S.C.
Institute of Molecular Medicine, R907
2121 W. Holcombe Blvd.,  Houston, TX  77030
713.500.2413-Voice//713.500.2424-FAX
-----------------
If everything seems to be going so well, you have obviously
overlooked something.
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