[Histonet] RE: human vimentin IHC

[Rene J Buesa]

Jan: 
I enjoyed your posting and I commend you for it.
This is what good professionals do, to share experiences and knowledge and this is one of the objectives of HistoNet.
René J.

--- On Wed, 7/15/09, Jan Shivers <shive003 <@t> umn.edu> wrote:


From: Jan Shivers <shive003 <@t> umn.edu>
Subject: Re: [Histonet] RE: human vimentin IHC
To: "histonet" <histonet <@t> lists.utsouthwestern.edu>, "PALMER Jason (SVHM)" <Jason.PALMER <@t> svhm.org.au>
Date: Wednesday, July 15, 2009, 10:18 AM


Just a comment from the veterinary world... in my validation tests of Vimentin (clone V9 by Dako), I found that xVimentin DOES cross-react with dog, cat, pig, cow, horse, donkey, goat, sheep, deer, chicken, rabbit, frog (weakly), and primate.  Dako's data from the past stated that V9 Vimentin did cross-react with hamster and rat, but NOT with mouse.  I have not tried it myself on these last three animals.

So, if Dako's data is accurate, and as long as you're only working with mouse tissue as your host stromal tissue, you should be OK in using Vimentin, at least Dako's V9 Vimentin.  But be sure to do your own validation on the separate species first to prove that your antibody does not bind to mouse tissue, before proceeding with the whole project.

I also use heat retrieval (pressure cooker) in Dako Target Retrieval Solution for Vimentin antigen unmasking.  Citrate buffer for antigen retrieval also works fine.

Jan Shivers
Univ. of Minnesota
Veterinary Diagnostic Laboratory
St. Paul, MN


----- Original Message ----- From: "PALMER Jason (SVHM)" <Jason.PALMER <@t> svhm.org.au>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, July 14, 2009 9:33 PM
Subject: [Histonet] RE: human vimentin IHC


Igor.

A few years back I used Dako V9 mouse anti human vimentin to label human grafts in a mouse background.  Tested it first on several mouse tissues and got no reactivity, compared to very strong reactivity in a variety of cell types in human tissue, and so am sure that it is human specific cf mouse.  I used the Dako ARK to get around the mouse-on-mouse background issues and was happy with the staining obtained (although not quite as sensitive perhaps as a standard, LSAB method).  I am actually about to try this again myself very soon.  I used citrate AR and primary at 1:800 for my staining.

Cheers,

Jason Palmer
Histology Laboratory Coordinator
Bernard O'Brien Institute
42 Fitzroy St, Fitzroy Victoria 3065
Australia
tel +61 3 9288 4018
fax +61 3 9416 0926
email: jason.palmer <@t> svhm.org.au


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

Message: 4
Date: Tue, 14 Jul 2009 15:03:47 -0400
From: Igor Deyneko <igor.deyneko <@t> gmail.com>
Subject: [Histonet] Human VIMENTIN and SMA IHC
To: Histonet <@t> lists.utsouthwestern.edu
Message-ID:
       <35e16a770907141203h14ccc18bt2e3123d11c478182 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Dear Histonetters!
I am wondering if anyone can possibly advise good antibodies for HUMAN anti
alpha SMA and Vimentin. I'm working with xenografts, human tumors with mouse
stroma and in the past had a lot of cross reactivity and background issues.
Does anyone know good antibodies or a clone, or has a good protocol for
either???
All would be greatly appreciated.
Thank you.
Igor Deyneko
Infinity Pharmaceuticals
Cambridge, MA

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