[Histonet] Frozens and antigen retrieval

[Lee & Peggy Wenk]

Usually not.

Fixation cross-links the proteins, which can mask the epitope (=antibody 
binding site of the antigen). So antigen retrieval breaks the fixative 
cross-links, exposing the epitope.

If there's no fixation, there's no cross-links, so the epitope is usually 
exposed and available to easily bind to the antibody.

Plus, there's no destruction of tissue morphology if you're not using 
antigen retrieval, so the quality of the section looks much nicer.

That being said, there may be some antibody out there that still needs 
antigen retrieval on frozen section, but then the company's protocol 
probably wouldn't say "optional".

So try it the first time without antigen retrieval.

Peggy A. Wenk, HTL(ASCP)SLS
Schools of Histotechnology
William Beaumont Hospital
Royal Oak, MI 48073

The opinions expressed are mine, and do not reflect Beaumont.
-----Original Message----- 
From: Daniela Bodemer
Sent: Wednesday, July 04, 2012 7:02 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Frozens and antigen retrieval

Hi all,

A question from my student: antigen retrieval when using frozens for 
immunofluorescence -yes or no?

The protocol suggested by the antibody company lists the retrieval as an 
option. I am used to do retrieval on paraffin sections, but not on cryo 
sections.

Hit me with your opinions on this :-)

Thanks in advance,

Daniela

Sent from my iPad

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