RE: Pancreatic Amylase

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From:"Hewlett Bryan (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA>, "''" <>
Date:Sun, 19 Sep 1999 09:40:56 -0400


We use the monoclonal anti- Pancreatic Amylase Cat# MU099-UC from Biogenex. 
Formaldehyde-fixed(16-24hrs) paraffin-embedded, 4 micron thick sections are
dried overnight at 45C, de-waxed in Xylene 3X10 min., rinsed X2 in 100%
ETOH, endogenous peroxidase blocked in 5% H2O2 in methanol for 30 min., 95%
ETOH X2 @ 1min, 70%ETOH-water. Sections are pretreated by digestion for 10
min@RT in 0.6% w/v Ficin dissolved in 0.01M NaOH containing 0.1mg CaCl2 per
100ml, pH is adjusted to pH7.8 with 1/N HCl. Following blocking in 1% v/v
Normal Goat Serum (NGS) in TBS, sections are incubated for 1 hour at RT in
1:100 dilution of Ab in 1%NGS in TBS. Detection is with LSAB(Zymed's
Histostain, GaM), chromogen is AEC.  The results have always been consistent
on our controls( pancreas fixed in NBF for several time points), test
specimens are prone to reflect the quality of fixation and processing(
particularly short fixation!).

Hope this helps

> ----------
> From:[]
> Sent: 	September 17, 1999 7:23 PM
> To:
> Subject: 	Pancreatic Amylase
> Does anyone out there have a pancreatic amylase that works on paraffin 
> embedded tissue?  We've tried and can't seem to get it to work.  Your help
> is 
> greatly appreciated.
> Thanks!
> Fran
> Yale University

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