I need to prepare a-amylase solution of concentration 2units/ml for an invitro a-amylase inhibition assay.
The a-amylase container I have does not mention the concentration of the enzyme and is in powder form. The following details are mentioned on the container:
Art No:K01042 (Kemphasol company) - Enclosed picture of label
CAS No: 9000-92-4
Amylase activity: 1:2000
I am not able to figure out how to prepare 2units/ml of a-amylase solution from a-amylase powder I have. Can someone help with any equation to convert units/ml to mg/ml
Thanks in advance
Namratha
Which company (for K01042 I find nothing in Google) and what are you buying, i.e. what it is written in the container. Perhaps a photo would be useful. In other words with the info you provide it is impossible. But with all the information it will be very easy to tell how to do it.
For isntance in Sigma you find ab enzyme from pig pancreas and they say ~1000 units/mg protein and there are 5 ml and 50 mg in the container. Then you have 1000 x 50= 50000 units in 5 ml. Then you have to calculate the amount needed for 2 units. Preferably in this case is first do one or two serial dilutions to have for instance 100 units/ml and then you just add the right amount to have 2 U/ml (the number of microliters will depend on the final volume used in your assay).
Rafael Franco , I have edited my question and included the label pictures as per your suggestion.
It says Amylase activity = 1:2000.
Do you know what this means?
One of my colleagues said it implies 1g contains 2000 U.
I am not sure if that is the correct interpretation as they have not mentioned any units on the label
1:2000 is not equivaleent to 1 g 2000 U (100% sure of it!!!)
I do not see images of label pictures
Impossible to know the activity of this powder with these lebels.
1. I hope this is pure as purity is not indicated.
2. I suggest you contact Kemphasol that seems is an Indian company. If they do not know the answer, at least ask from where amylase been prepared, from animal pancreas or from another source.
Then you can guess ging to SigmaAldrich page and see activity per mg if from pancreas (1000 units/mg), α-Amylase from Bacillus licheniformis (lyophilized powder, 500-1,500 units/mg protein, 93-100%)
3. Finally, please be aware that definition of unit for this enzyme according to Sigma is:
One unit will liberate 1.0 mg of maltose from starch in 3 min at pH 6.9 at 20 °C.
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