[Histonet] IHC validation

[Rene J Buesa]

I had never done that and it would be necessary only if the tissues you are using were fixed in a different way to yours.
Tissue fixation is the step that could affect reactivity because if the tissue has been fixed in an alcoholic fixative it would not need HIER and it is uncertain how this unnecessary step (in that specific case) could affect reactivity.
No other step (dehydration and clearing) could really affect the reactivity. Even temperatures above 45ºC have been demonstrated not to affect the epitopes.
Xylene do have an extracting effect on epitopes and it would be nice to know if the clearing agent was xylene.
Now, if you fix with NBF, dehydrate with ethanol and clear with xylene and the tissue you have from outside has been processed with a similar protocol, you do not need to validate that tissue. At least is how I see it.
René J.

--- On Thu, 9/1/11, Tim Higgins <thiggins <@t> cddmedical.com> wrote:


From: Tim Higgins <thiggins <@t> cddmedical.com>
Subject: [Histonet] IHC validation
To: histonet <@t> lists.utsouthwestern.edu
Date: Thursday, September 1, 2011, 12:22 PM


Hello Histo Friends,

Has anyone had to validate IHC antibodies when you do not have tissue processed in house to use for your validation.  I do have control tissue that stains properly but it was acquired from outsides sources with verified proper predicted staining.

If you have done this type of validation, how did you make the pathologist feel comfortable with it?  I understand pathologist all to well and you can't make them do anything they do not want to do but I am looking for suggestions that might have worked in helping them feel comfortable.  

Any information would be appreciated.

Thanks,

Tim
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