As Jennifer mentioned, enzymes have optimal temperatures at which to work. But also optimal pH. Is your 0.5% amylase in water or a correct buffer? Ray, Seattle, WA
----- Original Message ----- From: "Loree via Histonet Lager" <histonet@lists.utsouthwestern.edu> To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> Sent: Tuesday, November 3, 2015 12:37:04 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 CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
