[Histonet] Pas Digestion
Lager, Loree
loree.lager at seattlechildrens.org
Tue Nov 3 14:37:04 CST 2015
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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