[Histonet] RE: immunohistochemistry help please

[Sebree Linda A]

To test your deparaffinization Rebecca, I'd try staining a slide with something cheaper and quicker, like just your counterstain to see if that's working.  Then go from there.

Linda A. Sebree 
University of Wisconsin Hospital & Clinics 
IHC/ISH Laboratory 
600 Highland Ave. 
Madison, WI 53792 
(608)265-6596 
FAX: (608)262-7174 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Tarantelli, Rebecca Anne
Sent: Thursday, August 01, 2013 2:20 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] immunohistochemistry help please

Hi all,

I am new to this forum, but a friend recommended I send my issue out to you for help. Four weeks ago I completed a new protocol for Immunohistochemistry (IHC) using protinase K epitope retrieval, and monoclonal antibody 3F6. It worked perfectly. Three days later, after 13 beautiful stains, it just stopped working. The slides will not even take up the counter stain - Hematoxylin. I have exchanged my xylene, and tried again, and it still is not working. I am guessing since my counter stain isn't working, this is an issue in the deparaffinization stage, but I'm just not sure.

Please let me know if you have any suggestions!

Thank you
Rebecca

Rebecca Tarantelli

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