[Protein-analysis] Re: Formalin-fixation after snap freezing
Kyle Legate
via proteins%40net.bio.net
(by legatek from hotmail.com)
Tue Aug 28 14:23:59 EST 2007
RWILLDMD from aol.com wrote:
>
> My question: Is it possible to fix the previously frozen samples in
> formalin and continue analysis via immunohistochemistry for the protein
> (specifically proinflammatory cytokine) expression? Also, does minus 20 storage
> (instead of minus 80) put the tissue at risk?
>
I'm assuming you plan to section the tissue for IH studies. In this case
I would embed the tissue, unfixed, directly in cryomatrix, cut in a
cryotome, and fix them on the slide as you need them with 4% PFA on ice
for 10-15 minutes. I have not had any problems in the past by storing
blocks at -20 for a couple of months but it really depends on what
you're trying to detect. Trial and error is the only way to know.
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