Troutman, Kenneth A
kenneth.a.troutman <@t> Vanderbilt.Edu
Fri Jun 5 16:03:07 CDT 2009
I would recommend the following:
Citrate for 35 min at 95 deg. Cool down for 10 min. (This is our protocol.)
Be sure you are using charged slides and I would let them air dry for at least 1 hour before heating and deparaffinizing. That might help, too. Unfortunately, I don't think there is an effective method of retrieival that will completely eliminate the possibilty of floating tissue.
Ashley Troutman BS, HT(ASCP)QIHC
Department of Pathology
Vanderbilt University Medical Center
<http://www.vanderbilthealth.com/main/> <http://www.vanderbilt.edu/> <http://www.mc.vanderbilt.edu/>
Date: Thu, 4 Jun 2009 20:01:50 -0700 (PDT)
From: Hatem Salim <dr.hatemsaied <@t> yahoo.com>
Subject: [Histonet] immunohistochemistry
To: histonet <@t> lists.utsouthwestern.edu
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I am Research assistant at Physiology department of Michigan state university. I am using the (ß-Catenin Antibody (Carboxy-terminal Antigen) #9587 for IHC on mice femur sections. I have used this antigen unmasking method: For Citrate: Bring slides to a boil in 10 mM sodium citrate buffer pH 6.0 then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench top for 30 minutes.
The problem is that bone detachment always occurs so I wonder if there is a method of retrieval that prevents bone detachment.
> Waiting to hear from you soon.
> Thank you for your consideration
> Best wishes
> Hatem Salim
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