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 <[email protected]> wrote: From: Jan Shivers <[email protected]> Subject: Re: [Histonet] RE: human vimentin IHC To: "histonet" <[email protected]>, "PALMER Jason (SVHM)" <[email protected]> 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)" <[email protected]> To: <[email protected]> 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: [email protected] ------------------------------ Message: 4 Date: Tue, 14 Jul 2009 15:03:47 -0400 From: Igor Deyneko <[email protected]> Subject: [Histonet] Human VIMENTIN and SMA IHC To: [email protected] Message-ID: <[email protected]> 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 Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
