re: CD 34 Immunostain and Decalcification
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From: | "Michelle M. Sutorik" <istsmmmc@umich.edu> |
To: | HistoNet Server <HistoNet@Pathology.swmed.edu> |
Reply-To: | |
Date: | Fri, 04 Jun 1999 13:08:30 -0400 |
Content-Type: | text/plain; charset="us-ascii" |
Hello Everyone,
I have a question concerning a CD 34 stain. I am trying to stain
endothelial cells that are contained in a implant consisting of a Collagen
and hydroxyappetite carrier. These specimens are about an eighth of an inch
in diameter. They were fixed in 4% paraformaldehyde for 24 hours and
Decalcified for 2 wks in .5M EDTA, pH=8. which as I understand it is less
harsh than the acid decalcification technigues. I have obtained successful
staining of CD34 in a resorbable carrier that does not require
decalcification but so far have been unable to retrieve this antigen in the
presence of hydroxyappetite. I am wondering if and how decalcification
effects the antigen retrieval and also how to calibrate the decalcification
without overdoing it. Any ideas would be greatly appreciated.
I have tried using Microwave Antigen retrieval and also trypsin retrieval.
If anyone has had success retrieving this antigen in a boney tissue that
information would likely be helpful also.
Thanks Much,
Michelle M. Sutorik
Univ. of MIch.
Dept of Oral Pathology
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