re: CD 34 Immunostain and Decalcification

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From:"Michelle M. Sutorik" <>
To:HistoNet Server <>
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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