[Histonet] RE:Antigen retrieval paraffin skin samples

[Sally Price]

Maybe I've misinterpreted Nick's posting, but I think what he's looking for
is what kinds of reagents are used for retrieving Ck antigens.  It may
something to do with the PFA fixation, but I'm not too sure about using
ficin for CKs,  We usually use other enzymes or heat.  Proteinase K, or
trypsin, or citrate solutions work really well in most cases.
Sally
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Message: 8
Date: Thu, 30 Jul 2009 09:30:37 -0600
From: "Sorenson, Jon (Nampa)" <JonSorenson <@t> chiwest.com>
Subject: [Histonet] RE:Antigen retrieval paraffin skin samples
To: histonet <@t> lists.utsouthwestern.edu

We use and like the PASCAL pressure cooker system from DAKO, it is uniform,
convenient and fairly Easy to use.
Jon Sorenson
Histology Coordinator
Mercy Medical Center
jonsorenso <@t> chiwest.com
208-463-5267

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Message: 17
Date: Wed, 29 Jul 2009 14:05:29 -0700 (PDT)
From: "Nicholas David Evans" <ndevans <@t> stanford.edu>
Subject: [HISTONET] Antigen retrieval paraffin skin samples
To: <histonet <@t> lists.utsouthwestern.edu>

Dear all,
Might anyone be able to offer their advice on the best method to retrieve
antigens on paraffin embedded skin sections? I am immunostaining for
epithelial cytokeratins, and I have block-to-block variation in the quality
of the staining - some block give consistently good staining while others
giving negligible staining. The tissues were prepared by fixation in 4% PFA
for 24 - 48 hours followed by dehydration and embedding. I
currently use 15 mins Ficin digestion. I suspect that the differences may be
accounted for by variability in the length of time samples were fixed in PFA
and then 70% ethanol.
Best wishes
Nick