[Histonet] PAS with diastase

[Bryan Llewellyn]

I had the same problem some years ago.  I traced it to the malt 
diastase.  I switched to hog pancreatic alpha amylase from Sigma 
(product # A3176)and the problem immediately disappeared.  I used enough 
powder to cover a 1 cm circle about 1 mm deep dissolved in 15 mL (a test 
tube full) of distilled water and applied at room temperature for 30 
minutes.  That's not histochemically precise, but plenty good enough to 
remove all glycogen so the underlying PAS positive material can be 
demonstrated.  Buffers and increased temperature should increase the 
efficiency.

Bryan Llewellyn


MICHELE M wrote:
> Hi All,
>  
> We're looking for some help with our PAS with diastase stain.  For some reason, we can not get it to work. The PAS works fine but the diastase does not seem to be working.  We are using diastase of malt from Fisher and a 0.1M phosphate buffer at pH 6.0 from Polyscientific.  Our tech also tried using distilled water with the diastase instead of the buffer. The diastase solution is heated in a waterbath at 37 degrees for one hour. We have used liver, colon and esophagus tissue controls.  If anyone has any ideas what might be the problem, please let us know.
>  
> Thanks,
>  
> Michele
> 
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