Annexin V-FITC/TUNEL

Tom Mc Closkey thomasm at nshs.edu
Thu Oct 29 15:05:41 EST 1998



--- On Wed, 28 Oct 1998 23:21:34 -0500 (EST)  plett at auhs.edu wrote:

>
>Question: Since the Annexin-V stain should be run with a live/dead stain
>(PI or 7AAD), wouldn't fixing make all cells permeable to these stains?
>
>Any comments?


	I'm not sure that annexin "should" be run with a live/dead stain.  
In a well characterized system in which the cells are dying by apoptosis, 
the assessment of membrane integrity yields information aobout cells which 
are early [membrane intact] and cells which are further along [membrane 
disrupted].  If the goal of the assay is to quantitate total apoptotic 
cells, annexin alone is sufficient [provided you verify that the annexin 
positiv e peak represents apoptotic cells and is not some nonspecific 
interaction].  The omission of PI for membrane integrity also allows the 
possibility to examine phenotypic, cell cycle, or other parameters of 
interest.

Tom


--------------------------------------------------------
Thomas W. Mc Closkey, Ph. D.
Director, Flow Cytometry
North Shore University Hospital
Biomedical Research Center
350 Community Drive
Manhasset, Long Island, New York 11030
ph: 516-562-4844 [office]; 516-562-1135/4641 [lab]
10/29/98   12:15:50 PM
E-mail: thomasm at nshs.edu 
--------------------------------------------------------




More information about the Cytometry mailing list