I need to do McMannus' PAS stain. The protocol I have tells me to use 0.1-0.5% amylase solution in ddH2O (pre warned at 37 degrees for 1 hr). The problem is the prize. Amylase is too expensive for me to purchase. Is there any way I can substitute it with some other solution that is going to remove glycogen?
Dear Kathy,
despite faint smile at my thoughts... nevertheless not joking... and knowing the following might be an unscientific and/or unprofessional reply: since you cannot afford the commercially available (certified) amylase you can do it as in former centuries: try human (your own) saliva which contains amylase... not certified, not standardised, but nevertheless it should work ....
But I found also that it might be possible to substitute "amylase" by
As always with the value of postings in internet-fora: no warranty at all.
Best of luck, Wolfgang
Kathy,
I am working... had inserted my answer in this field, but forgot to klick to "Add your answer" before my refreshing the page...
It would last some hour to reconstruct my original reply.... but I would like to point you to the following (out of my memory):
If you are doing experimental staining I nevertheless would recommend to purchase "certified" amylase (indicating also the enzyme activity at a certain T) (google: ) even if it is "expensive" (not knowing whether you need alpha or ß-amylase).
3 other possibilities (substitutes):
i) your own (human) saliva contains amylase: "not certified", cheap, had been used and done in >>former times.
ii) I found also "raw honey"(contains natural plant amylases) and fruit to be perhaps a substitute for amylase enzyme,
iii) as well as (used as substitute for amylase in brewing beer and wine, also for bakery, perhaps can be purchased really cheap from local brewery shops) . Since the enzyme content will be low, you have to recalculate percentage valid for diastatic malt powder vs. amylase alpha or ß-enzyme activity.
[eventually look also for
Hope this helps at least for the first second.... best of luck, Wolfgang
Kathy, I am sorry...
it is a little bit intriguing for me...but some hour ago, when I wrote my "second" reply again I was not able to find my real first answer despite closing and reopening the threadpage... (I faced this behaviour of the reply-form sometimes already). So now you have both versions...any other reader: apologize for that circumstance.
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