Re: BrdU

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From:tylee <tylee@itis.com> (by way of histonet)
To:histonet <histonet@magicnet.net>
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Sharon,
Are you ready for some speculation??? I am not currently a hands-on user of
anti-BrdU.

I have looked into the various anti-BrdU clones in a past life and from
what I know,
all of the anti-BrdU antibodies require denaturation of the DNA to the
single stranded form to be able to recognize the incorporated modified
nucleotide. I suspect that one problem could be the differential ability to
denature the DNA in the two tissue types. From what I remember, acid, heat,
or nucleases are the most common methods to denature the DNA. I do not
specifically recall which clones Amersham and BM have (or have licensed);
but there are numberous possibilities out there that work well.

Another possibility could be the differential penetration of antibodies to
the nucleus of the two different tissues.

The rate of proliferation may have an affect on the rate of development if
the number of antigenic sites is the limiting factor in development;
however, at 1-2 or 10-15 minutes, I doubt if that is the case.

My two cents worth.

Ty Lee

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From: sbledsoe <sbledsoe@iupui.edu>
To: HistoNet@Pathology.swmed.edu
Subject: BrdU
Date: Wednesday, February 03, 1999 9:28 AM

I would appreciate any comments that I can pass along to the higher powers
that be....

I have been staining vessels and intestine with BrdU (Primary is either
Amersham or Boehringer Mannheim, secondary is Horse-anti-mouse-botin rat
affinity purified, followed by Vector ABC kit then DAB).... the intestines
stains very nicely within 1-2 minutes (with no background) while the
vessels take 10-15 minutes.  I have been asked why and I have no idea.  Why
would a nuclei of the intestine stain much faster than a nuclei of a
vessels.  The vessels are rat arteries that have been subjected to
pressure, so they do have a lot of cell proliferation.

Thanks
Sharon




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