Re: CD34 measurement

Dennis_Young@CIS.ucsd.edu
Thu, 26 Oct 1995 08:55:00 -0700

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Maybe in reply to: Ian Titley Leukaemia Reaserch Fund Centre: "CD34 measurement"

As of right now, we use BD's Leukogate (CD45=LCA/CD14), isotype controls, and
anti-CD34 (HPCA-2) in separate tubes (FACScan). We estimate %CD34 by correcting
the %+ in the CD34 tube (by counting low SSC vs. CD34) with the %CD45+.

(CD34 - Control)/CD45
HPCA-2 binds the protein backbone of CD34; we use this method to stain
peripheral blood, leukapheresis product, bone marrow, umbilical cord blood, and
chymopapain-treated, enriched cells.
The Leukogate allows us to also estimate %CD34 as a fraction of mononuclear
cells. We also calculate CD34/ul based on WBC.

We plan to phase in a CD45/CD34 method soon.

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* Dennis J. Young Voice : (619) 822-0407 *
* Flow Cytometry Core Facility FAX : (619) 822-0412 *
* University of California, San Diego USA e-mail: djyoung@ucsd.edu *
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______________________________ Reply Separator _________________________________
Subject: CD34 measurement
Author: ian_t@icr.ac.uk at @UCSD
Date: 10/25/95 3:27 PM

Further to Ed's request about how people measure CD34+ cell numbers by flow.
As well as asking about whole blood vs mononuclear preps and whether to use
CD45 or not (and indeed which CD45 antibody) it would also be interesting to
know which clone of CD34 antibody people use (since there are at least three
distinct epitopes on the CD34 molecule).

Thanks

Ian Titley
Leukaemia Research Fund Centre
at the Institute of Cancer Research
237 Fulham Road
London

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>Subject: CD34 measurement
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