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@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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