[Histonet] Pepsin
James L Burchette
burch007 <@t> mc.duke.edu
Fri Oct 10 12:39:00 CDT 2008
Gene:
Dako and Biocare both sell the stable pepsin solution. It is easy to
prepare if you want to make it yourself.
I use this pepsin everyday. Here is the recipe that Dr. Brigati gave me.
Best Regards,
Jim Burchette
Brigati’s 0.25% Stable Pepsin Solution
Reagents
Pepsin, #P7012, Sigma Chemical Co.
** 10X Automation Buffer, #BMM30, Fisher Scientific **
Preparation
40 ml 10X Automation Buffer
356 ml distilled water
Adjust mixture of Automation Buffer and water to pH 2.0.
Add 1.0 gram of P7012 pepsin (it is important to add the pepsin after the
pH adjustment). Place container on a stir plate with gentle stirring
allowing the pepsin to dissolve.
The solution is sometimes cloudy at first, but will become clear after
sitting overnight.
Store at 4 C. Apply at room temperature
Digest formalin-fixed, paraffin embedded tissue sections for 13-15 minutes
at 37-40.
Wash with IHC procedural buffer or DI water to stop enzyme digestion and
to neutralize the pH.
** The Biomeda TBS automation buffer is no longer available. I now use the
Automation Wash TBS Buffer from Biocare Medical. It is a 20X concentrate.
Adjust the volume of water and buffer accordingly. You could also use 396
ml of 1X TBS.
Dr. Brigati passed away a year ago. He was a great pathologist, mentor and
friend.
njoydobro <@t> aol.com
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
10/10/2008 09:05 AM
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Histonet <@t> lists.utsouthwestern.edu
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Subject
[Histonet] Pepsin
Does anyone have a catalogue # for the Pepsin used in Brigati's stable
pepsin?
Thanks,
Gene
Cleveland Clinic
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