RE: [Histonet] MGMT antibody

From:"Favara, Cynthia \(NIH/NIAID\) [E]"

I am not familiar with this antibody but I use the Ventana system with a
number of research antibodies and find that I need to boost the
Streptavidin often with antibodies not provided by Ventana. You may not
be able to do this if you do not have their research software. In that
case I would try an overnight incubation.

Hope this is helpful - if you would like more specifics about the boost
let me know.

c 

Cynthia Favara
NIAID/NIH/RML/LPVD
903 South 4th Street
Hamilton, MT 59840
406-363-9317

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-----Original Message-----
From: Elaine Dooley [mailto:DOOLEEO@shands.ufl.edu] 
Sent: Thursday, May 11, 2006 10:08 AM
To: Histonet@pathology.swmed.edu
Subject: [Histonet] MGMT antibody

Dear Histonetters,

I have been trying to get MGMT antibody to work on a Ventana Benchmark
immunostainer and I am not getting the good results yet.  The MGMT
(clone MT 3.1 from Novus Biologicals) supposed to stain Colon carcinoma.
 So I tried 1:100 with CC1 antigen retrieval and did not get any
staining.  I tried 1:20 with amplify and CC1 retrieval and everything
was staining.  So I tried 1:40 with and without amplifier,  both with
CC1 retrieval and they stained everything weakly.  Perhaps a different
retrieval should be used. Or maybe it does not stain all colon
carcinomas and I am trying to make one stain that really should not.  I
have limited time and thought if anyone might have some insight I would
appreciate it.

Elaine Dooley
HTL
Shands labs at Rocky Point
352-265-0111 ext 72117

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