RE: eNOS for paraffin-embedded sections??

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From:Todd Sherman <tsherman@flash.net>
To:histonet@pathology.swmed.edu
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Hello Kathie,

Although I have left the world of research IHC, I worked with anti-eNOS
antibody from Transduction Laboratories with great success for several years
at UT Southwestern Medical Center at Dallas.  We performed highly specific
staining in formalin-fixed, paraformaldehyde-fixed and frozen sections using
the monoclonal Ms IgG (product #N30020) in rat, sheep, and human tissue.
Granted, your mileage may vary...but the pertinent factors for obtaining the
best signal in aldehyde-fixed, paraffin-embedded specimens were antigen
retrieval in basic buffer (we used AR10 from Biogenex w/ microwave HIER),
not baking sections to slides (air-dry overnight at RT only), and overnight
incubations with the primary at 4 deg Celsius.  Baking the sections will
certainly destroy this epitope.  This antibody was not cross-reactive with
the other constitutive isoform ,nNOS, in our hands.

see: www.translab.com/DS/Pages/N30020.html

Hope this helps,
Todd Sherman

-----Original Message-----
From: Kathie Berghorn [mailto:kab35@cornell.edu]
Sent: Monday, July 31, 2000 2:18 PM
To: histonet@pathology.swmed.edu
Subject: eNOS for paraffin-embedded sections??


Hi,
	I am in search of a primary antibody for endothelial nitric
oxide synthase (eNOS) antibody that will work in paraffin-embedded
sections.  I have searched Sciquest and talked to Zymed, Stressgen
and Sigma-Aldrich (RBI) without success.

	Should anyone have information, please contact me.

	Thanks much,
	Kathie







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