[Histonet] RE:Antigen retrieval paraffin skin samples

[Sorenson, Jon (Nampa)]

We use and like the PASCAL pressure cooker system from DAKO, it is
uniform, convenient and fairly 
Easy to use.
Jon


Jon Sorenson
Histology Coordinator
Mercy Medical Center
jonsorenso <@t> chiwest.com
208-463-5267

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>
Message-ID: <002c01ca1090$4d32f280$ce1d41ab <@t> DellDesktop2>
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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