Diastase powder will dissolve in any water, but requires some calcium ions to function. Thus it will not perform in distilled water, but will perform well using plain tap water. Also, the diastase powder from sigma will work well if only a very small amount of the powder is used. We always used just enough to barely cover the bottom of a glass coplin jar and then fill with tap water. I would expect 0.5 grams in 50 mls of water is probably too much. Try using one tenth that amount or 0.05 gm in 50 ml. When there is too much diastase in solution, enzyme conformation is inhibited and it does not work. Spokane water is quite hard. It always worked well at room temperature for 20 minutes.
Mickie Mickie Johnson, B.S., HTL(ASCP) Mohs Histology Consulting Services, LLC & Mohs Lab Staffing 2507 S. Manito Blvd. Spokane, WA 99203 509-954-7134 FAX 509-624-3926 Web: www.mohshistogyconsulting.com & www.mohslabstaffing.com Email: [EMAIL PROTECTED] DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. -----Original Message----- From: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Rene J Buesa Sent: Friday, October 24, 2008 8:38 AM To: histonet@lists.utsouthwestern.edu; McKnight, Tanisha Subject: Re: [Histonet] PAS with Glycogen Digestion - Solution Not Working Tanisha: Being an enzyme, you cannot use diastase in distilled water. You have to prepare a diastase buffer at pH7 to dissolve the diastase and do the digestion at 37ºC during 30 minutes. That will work. Please, do not try to use saliva! René J. --- On Fri, 10/24/08, McKnight, Tanisha <[EMAIL PROTECTED]> wrote: From: McKnight, Tanisha <[EMAIL PROTECTED]> Subject: [Histonet] PAS with Glycogen Digestion - Solution Not Working To: histonet@lists.utsouthwestern.edu Date: Friday, October 24, 2008, 11:26 AM Hi All: I am in the process of working up a PAS Staining protocol. The PAS stain is working fine, but I am unable to get my tissue to digest. I am using .5g of Diastase in 50ml of DI water. I tried heating the solution in a water bath for 30 min, 1 hour and 1.5 hours and it still looks the same as the one without digestion. Could I be using the wrong Diastase? If so, does anyone have any suggestions for a new product? Thanks, Tanisha Neely, HT(ASCP) AP-Histology/Specimen Management <mailto:[EMAIL PROTECTED]> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet No virus found in this incoming message. Checked by AVG - http://www.avg.com Version: 8.0.175 / Virus Database: 270.8.3/1747 - Release Date: 10/26/2008 9:27 AM _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet