Antigen Retrieval

From:Amos Brooks

    Generally it is a bad idea to try to get all your antibodies to be run
the same. It just doesn't work that way. Most cytokeratins prefer enzymes.
Actually using HIER (heat induced epitope retrieval) on AE1/AE3 cytokeratin
cocktail causes PMN (polymorphonuclear) cells of bone marrow and blood to
stain. Hematopathologists will hate you for it ... Yes, I did find this out
by personal experience. My ears are still ringing from a year ago!
    Since it is cheaper and easier, always try to get the antibody to stain
with no retrieval at all. Often, this just won't work. so try HIER. It is
wise to compare this to using an enzyme. You'll get much better results if
you tweak your protocols with some experimentation.
    Also, remember to read your spec sheets. The manufacturers usually test
their products on plain old formalin. Changing anything will throw curve
balls in procedure development.

Amos Brooks
Laurie Colbert wrote:

  I was wondering if it was necessary to use zinc formalin to enhance IHC
staining and prevent cross-linking if you were doing antigen retrieval for
every antibody.  Any comments???
  Laurie Colbert

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