BrdU
ronald hill (ronald@sugen.sf.ca.us)
Mon, 30 Sep 1996 09:42:24 -0700
Larry Seamer (CYTOM) wrote:
>
> I can provide some information concerning the potential pitfalls part of
> your question by refering you to a paper by Cinthia Hoy and myself from a
> few years ago:
>
> Thermal Denaturation of DNA for Immunochemical Staining of Incorporated
> BrdUrd: Critical Factors That Affect the Amount of Fluorescence and the
> Shape of BrdUrd/DNA Histogram: Cytometry 10:718-725 (1989).
>
> On -1 xxx -1 SMTP%h82@aixterm1.urz.uni-heidelberg.de@COBRA.UNM.EDU wrote:
>
> > Is anybody of you aware of a good (means working) protocol for
> > a Br-desoxyuridine-FACS-procedure? I have used the B-D-protocol
> > provided in their monoclonal antibody resource book, but without
> > success :-(
> > In particular I am looking for information about the FACS-setup
> > for aquisition and analysis of cell proliferation and hints
> > about possible pitfalls in the procedure.
> >
> > Any help is greatly appreciated.
> >
> >
> > --
> > Andreas Schr=F6der
> >
> >
> >
I recently posed a similar question to the list. Denaturation of the
DNA turned out to be the critical variable in my hands also. Thermal
denaturation resulted in a detectable signal when acid denaturation
did not. However, I also received a recommendation to use more
aggressive acid denaturation. But I have not tested this yet.
-Ron Hill, Ph.D.
Research Scientist
Sugen, Inc.
Redwood City, CA
ronald@sugen.sf.ca.us
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