[Histonet] IHC Staining Problem

[Ross Benik]

We have developed a staining protocol for the CD79a antibody from two
different vendors and both require a HIER TRIS/EDTA pH 9.0 retrieval. 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Amos
Brooks
Sent: Monday, November 01, 2010 2:34 PM
To: jhill <@t> vet.k-state.edu; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] IHC Staining Problem

Hi,
     I think you mentioned using citrate retrieval. Have you tried high
pH
retrieval with EDTA or an equivalent? I think CD79 works better with
high pH
retrieval.

Amos


Message: 1
Date: Sun, 31 Oct 2010 19:29:59 +0000
From: Jennifer Hill <jhill <@t> vet.k-state.edu>
Subject: RE: [Histonet] IHC Staining Problem
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
 
<8AA2173DC209CA438077A832FF98BD7F02806542 <@t> VETMXHT.ads.vet.k-state.edu
>
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Thanks to everyone who has responded so far.  To answer the questions
asked,
our controls are either being cut the same day or the day or two before,
so
they are not setting for extended periods.  We have also used at least
three
or four different controls, all with the same results. (The controls
stain
fine for other tests: CD20, CD3, etc.)The pathologists are saying that
the
patient slides are not staining as well.  So it doesn't seem to be
something
so simple as a control issue.  As to the dehydration/deparafinization,
that
takes place on an automated stainer, and no other stains are having
issues
so I don't believe that is the issue either.


Jennifer Hill
Kansas State University
Veterinary Diagnostic Lab
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