[Histonet] Microwave Processing

[Margaret Blount]

Just a quick suggestion which maybe you have already tried. Most alkaline
phosphatase commercial substrates use Levamisole as an inhibitor of
endogenous alkaline phosphatase. This does not inhibit the intestinal enzyme
as used for labelling antibodies, so maybe that is what you need. The method
was published by Ponder and Wilkinson, but I don't have my notes to hand so
can't give you the exact reference, however if you look in any reputable
textbook on immunohistochemistry you should find the details there.

margaret

Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Maria
Limberis
Sent: Wednesday, April 21, 2004 5:34 PM
To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson;
Steven E. Slap; kevin williams; histonet <@t> pathology.swmed.edu
Subject: RE: [Histonet] Microwave Processing


Hi everyone-
I have instilled a vector expressing human placental Alkaline Phosphatase
in the lung of rhesus monkeys. However, when I use a protocol routinely
used in our lab to prevent background in mouse lung (which works very well)
the background is very high preventing assessment of gene transfer. I am
wondering if anyone has any suggestions or ideas!
Much appreciated
Maria

-------------------------------------------------------------------------
Maria Limberis, PhD
Gene Therapy Program
University of Pennsylvania School of Medicine
Department of Medicine
Division of Medical Genetics
The Wistar Institute, Room 201
3601 Spruce Street
Philadelphia, PA 19104

Tel: 215 8987166
Fax: 215 5737414
Email: limberis <@t> mail.med.upenn.edu


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