Re: [Histonet] PAS/D digestion problem

From:Rene J Buesa



John:
  I always used successfully the following procedure:
   
  a- prepare a "diastase buffer": anhydrous sodium chloride (8 g) + anhydrous dibasic sodium phosphate (0.282 g) + anhydrous monobasic sodium phosphate (1.974 g) + distilled water UP TO 1 liter. This buffer will have a pH @ 6 and has to be kept in the refrigerator.
   
  b- glycogen digesting solution: take 40 mL of the buffer and heat it to 37ºC and add 0.5 g of diastase of malt (Fisher brand "maltin" cat.No. D22-100).
   
  c- incubate the hydrated section at 37ºC for 30 minutes
   
  Proceed with the PAS, counter stain, etc., etc.
  René J.
  

Josh Wray  wrote:
  
 

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