[Histonet] Pepsin

[James L Burchette]

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 
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10/10/2008 09:05 AM

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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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