Inka, I use Immunon Trypsin Type II Digestive Enzyme Concentrate from
Shandon-Lipshaw (1-800-245-6212), catalog number 48444435.  It comes in
bottles of 20 tablets each, and 1 tablet is disolved in 25 ml of distilled
water (directions are enclosed with each bottle ordered).  I'm sure if you
called and asked for a copy of the product insert, they would be happy to
fax it to you.  After hydration, we place the slides in the trypsin solution
at room temperature for 15 minutes exactly.  Then they are rinsed with
distilled water and are ready for immunostaining (we have an automated
system).  Hope this helps.  Sue O'Brien
           Burdette Tomlin Memorial Hospital
           Cape May Court House, NJ  08210
           sobrien@bthosp.com

> ----------
> From: 	Jeff Silverman[SMTP:peptolab@hamptons.com]
> Sent: 	Friday, June 04, 1999 6:13 PM
> To: 	histonet@pathology.swmed.edu
> Subject: 	Re: trypsin treatment for overfixed tissue, CD31, Vendors
> need love too
> 
> To Inka Tertinegg,
> I use and recommend  the  prebuffered tablets sold by Sigma Chemical Co.
> I'm home now so don't have the catalog number, but Go to
> www.sigma.sial.com
> and order the prebuffered trypsin tablets. I keep them in the freezer and
> dissolve one or two for each run, it takes about five minutes to dissolve.
> They cost 20 dollars US for 50 I think. I use them for 30 minutes at 37
> degrees C for antigen retrieval for cytokeratin AE1/AE3, factor XIIIa,
> CD68
> and others. Very easy and reproducible. Good luck. 
> To Andrea at Imclone: I've always used either the trypsin 30 min at 37
> degrees C or citrate buffer in a microwave for five minutes on high and
> fifteen minutes on medium low  to detect CD31 using Biogenex Super
> Sensitive ready to use antibody with no problem. Antibody incubation is
> overnight at room temp. 
> To all my Histonet friends: Let's pray that a vendor's love letter somehow
> doesn't find its way onto the Histonet :>)  Have a happy.
> 
> Jeff Silverman
> Southampton Hospital
> Southampton NY USA
> 
> ----------
> > From: Inka Tertinegg <inka@playfair.utoronto.ca>
> > To: Histonet@pathology.swmed.edu
> > Subject: trypsin treatment for overfixed tissue
> > Date: Friday, June 04, 1999 5:02 PM
> > 
> > Hi,
> > 	I' recently seen a couple of references to trypsin pretreatment of
> > overfixed tissue. Would anyone have a protocol for this that they could
> > share?(ie. concentration of trypsin, buffer and time of treatment).
> > 
> > 	Thanks
> > 
> > 
> > 	
> > Inka Tertinegg
> > University of Toronto
> > (416)603-5800 ext 2850
> > inka@playfair.utoronto.ca
>