[Histonet] immunohistochemistry help please

[Rene J Buesa]

If the tissues you are trying to do IHC on are of different origin or processed differently that could be the cause.
I really advise you to do dewaxing the way "sophisticated and expensive instruments do" it is: use a 2% aqueous solution of dish washing soap at 90ºC for 1 minute, twice, then hot running water and then distilled water → to your IHC protocol.
This is the safest, cheapest, best and expeditious way of dewaxing sections.
René J.


________________________________
From: "Tarantelli, Rebecca Anne" <rat37 <@t> pitt.edu>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, August 1, 2013 3:20 PM
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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