[Histonet] immunohistochemistry

[Troutman, Kenneth A]

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.  
 
 Good luck.
 
Ashley Troutman BS, HT(ASCP)QIHC
Histopathology Laboratory
Department of Pathology
Vanderbilt University Medical Center
Nashville, TN
<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
Message-ID: <782588.78944.qm <@t> web46103.mail.sp1.yahoo.com>
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HI
 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
<http://www.vanderbilthealth.com/main/>