Re: Apoptosis in CD34+ cells

Ray Hicks (hicks@icrf.icnet.uk)
Tue, 30 May 1995 14:13:57 +0100 (BST)

You could try Annexin V, which picks up changes in membrane lipids
(specifically PS) which correlates well with other methods of apoptosis
detection. It works on intact unfixed cells and detects rearrangement of
the lipids before loss of membrane integrity. You should be able to
double label with annexin V-FITC and cd34-PE with little or no problem.

Ray

On Thu, 25 May 1995, STEVEN DAVID GORE wrote:

>
> We have been trying to adapt the TUNEL assay to identify apoptosis among
> normal human CD34+ cells, double labeled with PE-labeled CD34 antibody.
> While this procedure works well for CD34-expressing cell lines, such as
> TF-1 or KG1a (the CD34 labeling performed before formaldehyde fixation,
> then followed by MeOH or EtOH etc), among isolated bone marrow CD34+
> cells, CD34 appears undetectable after the alcohol second fixation step
> (either alcohol). Does anyone have any experience or suggestions?
>
>
>
> Steve Gore
> Johns Hopkins Oncology Center
> Oncology 2-109
> 600 N. Wolfe St.
> Balto, MD 21287-8963
> sdgore@welchlink.welch.jhu.edu
> 410-955-8781
> FAX 410-614-1005
>