[Histonet] RE: T Cell Heat retrieval using microwave pressure
I am using the rabbit anti-CD3 monoclonal antibody (SP7) from
LabVision at 1:500 (60 min, RT) visualized by EnVision anti-rabbit/HRP
and DAB+. HIER is done with Tris-EDTA pH9.0 (15 min boiling + 10 min
cool-down). In case you prefer citrate pH6.0 then go down to 1:200
with the primary.
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
phone: +31 20 5665631
Date: Wed, 15 Mar 2006 11:33:10 -0500
From: "Diana McCaig"
Subject: [Histonet] T Cell Heat retrieval using microwave pressure
To: "Histonet \(E-mail\)" email@example.com
I am trying to demonstrate T cells using Polyclonal Rabbit Anti human
CD3, T cell (DAKO ENVISION). The control tissue is tonsil and using a
manual procedure. I have increased the time in the microwave pressure
cooker from 5 up to 30 minutes (pH 6 buffer) and have no success. The
antibody is incubated for 30 minutes (recommended) but I have gone as
high as 1 hour. I am using AEC and Mayers counterstain. Any help or
advise would be greatly appreciated.
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