Re: CD10 antibody

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From:Kappeler Andreas <kappeler@patho.unibe.ch>
To:Histonet <HistoNet@pathology.swmed.edu>
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<!DOCTYPE HTML PUBLIC "-//W3C//DTD HTML 4.0 Transitional//EN"> <HTML><HEAD> <META content="text/html; charset=windows-1252" http-equiv=Content-Type> <META content="MSHTML 5.00.2314.1000" name=GENERATOR> <STYLE></STYLE> </HEAD> <BODY bgColor=#ffffff> <DIV>Hi Richard</DIV> <DIV>We are using Novocastra's mo-a-hu CD10, clone 56C6, Cat # NCL-CD10-270. Working conc. is 0.6 ug Ig/ml. Ask your supplier to specify the Ig concentration of the undiluted product  -  Novocastra has this information and provides it on request (that's at least something - some companies provide the Ig-conc of their product from the very beginning [that's where I like to shop], others will never give you this information [that's where I start to have some doubts about ... and prefer to go elsewhere]). Anyway, the antibody works reasonably well on formalin-fixed tissue, although overfixation is absolutely detrimental. We do a HIER using a microwave protocol and a 100 mM Tris buffer pH 9.5 (I know this is nonsense, but that's the way it 's published - and it works) with 5% urea (your tissue will not like the urea, but the staining will be better than after citrate buffer). Alternatively - and if you have money to burn - you can probably try one of the commercially available high pH antigen retrieval buffers.</DIV> <DIV>Good luck!</DIV> <DIV> </DIV> <DIV>Andi Kappeler<BR>Institute of Pathology </DIV> <DIV>University of Bern, Switzerland<BR></DIV></BODY></HTML>
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