Re: trypsin treatment for overfixed tissue, CD31, Vendors need love too

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From:Jeff Silverman <peptolab@hamptons.com>
To:histonet@Pathology.swmed.edu
Reply-To:
Date:Fri, 04 Jun 1999 18:13:24 -0400
Content-Type:text/plain; charset=ISO-8859-1

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



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