[Histonet] AE1/AE3 cross reactivity

[Mighnon Lashus]

We do not use AR for the AE1/AE3 on our Ventana system.  We use Protease 1 for 4 minutes.  I will check into the heat.  Any other suggestions?  We do the stains for Dr. Hessler.
Thanks,
Mighnon Lashus

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sally Price
Sent: Wednesday, January 16, 2008 5:53 PM
To: Richard Hessler, M.D.; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] AE1/AE3 cross reactivity

Dr. Hessler:
I have observed the phenomenon that you've described for other antibodies
more times than I can remember.  Personally, I believe that it has to do
with the way the Ventana system sprays on the retrieval solution
and heats slides during antibody/detection incubation periods, which is too
aggressive.  I think that an open system that doesn't heat slides or perform
AR isn't likely to produce these unexpalined reactions, but I digress.  So,
how do you fix the problem?  I say disable the on-line AR, and perform it in
a decloaking chamber, set at 100 degrees for 10 minutes.  Or, you could
consider using another pan-CK antibody, like Lu-5.
Good Luck,
Sally Price

----------------------------------------------------------------------------------------------

Date: Wed, 16 Jan 2008 07:57:41 -0600
From: "Richard Hessler, M.D." <rhessler <@t> pathgroup.com>
Subject: [Histonet] AE1/AE3 cross reactivity
To: "histonet <@t> lists.utsouthwestern.edu"
    <histonet <@t> lists.utsouthwestern.edu>
"
I have used this antibody for years but now have issues. Previously it was
the best choice for breast sentinel nodes, but now AE1/AE3  stains plasma
cells like CD138. It also stains a number of different sarcomas and
beautifully highlights reactive astrocytes. I had previously used Dako Ab
and autostainer with no problems. Now I have Ventanna and have noticed this
with both Ventana and Dako Abs used on that system.

Any thoughts???

Richard B Hessler, MD
Chief of Pathology
Erlanger Medical Center
Chattanooga, TN
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