[Histonet] Pas Digestion

[Jennifer MacDonald]

The optimal temperature for most enzymes in 37 degrees Celsius. 




From:   "Lager, Loree via Histonet" <histonet at lists.utsouthwestern.edu>
To:     "'histonet at lists.utsouthwestern.edu'" 
<histonet at lists.utsouthwestern.edu>
Date:   11/03/2015 12:38 PM
Subject:        [Histonet] Pas Digestion



Hello,

This is my first time using the histonet, as our veteran user retired.

We've been having an intermittent problem with incomplete glycogen 
digestion on PASD.  Our digestion solution is 0.5%  α-Amylase, Type VI-B 
from porcine pancreas (Sigma), which we freeze in small aliquots and thaw 
for 45 minutes at room temperature before use. We digest for 20 minutes @ 
room temperature, dropping amylase on slides.

In troubleshooting the issue, we discovered amylase was several years old, 
so new amylase ordered.   The new lot (while the same product number) 
produced even worse results.  After a closer look we saw that the new lot 
contained ½ of the units/mg solid, 26 to 13, not sure why?

Anyone willing to share their procedure or suggestions to help us resolve 
this issue?

Thank you,
Loree Lager
Seattle Children's Hospital
Histology




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