[Histonet] CD4/CD8

[Patsy Ruegg]

The problem I have found trying to do T subsets on mouse ffpe tissue
is that the antibodies are mouse monoclonals and so you must use a mouse on
mouse biotinylation detection system ( or at least I have not found a
non-biotin mouse on mouse detection, if there is one out there please let me
know) so with this biotin system you can get a lot of endogenous biotin
background even with serious AB block methods.
Patsy

-----Original Message-----
From: histonet-admin <@t> lists.utsouthwestern.edu
[mailto:histonet-admin <@t> lists.utsouthwestern.edu]On Behalf Of Amos Brooks
Sent: Thursday, November 06, 2003 11:04 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] CD4/CD8

Manuelle,
         I would certainly recommend using Envision (or some non-biotin
based detection system). Both antibodies work best using high pH retrieval
which tends to necessitate Envision.
         I can't speak for Dako CD4 as we use Novocastra but we get much
better results doing the peroxidase block prior to antigen retrieval. The
H2O2 interferes with the signal. It sounds like a pain but the results are
clearly better.
good luck,
Amos Brooks

At 11:35 AM 11/6/03, you wrote:
>Message: 19
>Reply-To: "Manuelle De Bock" <manuelle.debock <@t> UGent.be>
>From: "Manuelle De Bock" <manuelle.debock <@t> UGent.be>
>To: <histonet <@t> lists.utsouthwestern.edu>
>Date: Thu, 6 Nov 2003 11:36:03 +0100
>Subject: [Histonet] CD 4/CD 8 staining of frozen stomach sections of mice
>
>This is a multi-part message in MIME format.
>
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>Hello everyone,
>
>I am trying (for a long time allready) to produce a good CD4/CD8 =
>staining of frozen stomach tissue (mice). We use the Dako rat anti-mouse =
>CD4 antibody but always encounter a large amound of background  after =
>staining (it seems like there is a lot of cross reactivity with the =
>glandular epithelium). Can anyone help me to solve this problem ?
>Thanx !
>
>
>Manuelle De Bock, dierenarts (DVM)
>Laboratory of Veterinary Pathology=20
>Department of Pathology, Bacteriology and Avian Diseases
>Faculty of Veterinary Medicine
>Ghent University
>Salisburylaan 133
>B9820 Merelbeke - Belgium
>Tel 0032(0)9 264 7745
>Fax 0032(0)9 264 7789


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