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 ------------------------------
Message: 8 Date: Thu, 30 Jul 2009 09:30:37 -0600 From: "Sorenson, Jon (Nampa)" <jonsoren...@chiwest.com> Subject: [Histonet] RE:Antigen retrieval paraffin skin samples To: histonet@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 jonsore...@chiwest.com 208-463-5267 ------------------------------ Message: 17 Date: Wed, 29 Jul 2009 14:05:29 -0700 (PDT) From: "Nicholas David Evans" <ndev...@stanford.edu> Subject: [HISTONET] Antigen retrieval paraffin skin samples To: <histonet@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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet