RE: [Histonet] T Cell Heat retrieval using microwave pressure cooker

From:"Elizabeth Chlipala"


We used dako's ph9 retreival in the pascel with good results and
envision rabbit as a detection system.  We were staining human thymus
grafts in mouse tissue.  30 minute incubation at 1:100.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
[] On Behalf Of Diana
Sent: Wednesday, March 15, 2006 9:33 AM
To: Histonet (E-mail)
Subject: [Histonet] T Cell Heat retrieval using microwave pressure

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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