My stock solution is about 12000 U/mL
in relation to plant extract what concentration of alpha amylase is recommended and can represent the in vivo conditions ?
Thank you doctor Cheurfa
what i really need is to know if i can use 240 U/mL or it is better to use 1 U/mL or less
some papers used 1000 U/mL
is there any standarization ?
Dear Dr. Marmouzi,
Yes you are right, different article demonstrates alpha amylase inhibtion assay using different units of alpha amylase.
The basic principle as per sigma aldrich is:
One unit will liberate 1.0 mg of maltose from starch in 3 minutes at pH 6.9 at 20°C.
And as per sigma aldrich they are using 1 Unit/ml.
We personally use 8 Units/ml of alpha amylase, but I believe 240 U/ml will be too high.
The thumb rule here is you need to prepare all standard curve, proper controls i.e. positive control, negative control and in case of plant extract, sample control is must. You fix one concentration according to the generalized information availble for this assay, but need to stick with it throughout your study as well as need to mention it clearly while displaying and comparing your results with existing literature.
Dear Dr Rajeev K Singla
Thank you for your interesting response
How about studying the mechanistic way of inhibition
Can you recommend to me some good references for kinetic studies
Thank you very much
Dear Dr. Marmouzi,
I haven't came across any signifcant article which will provide details about the kinetic study during alpha amylase inhibition assay. we are trying to working it out.
Right now we are doing mechanistic analysis using docking studies.
Dear Dr Rajeev K Singla
It will be really interesting if we can collaborate in this direction
I am interested in Oat chemistry and bioactivities
and i would like to study some Oat compounds for their enzyme inhibition ability using docking models
If you are interested or you can help me
we can communicate via mails
Thank you again for your kind answers
Dear Dr. Marmouzi,
Thanks a lot for this opportunity. Let we discuss it on mail. Please drop me detailed mail at [email protected]
I must inform you Dr. Marmouzi that I am over engaged in multiple assignments as of now. So if at all we have any collaborative work together, then also it will be a time taking. I can't do any work instantly rightnow.
Regards
RKS
Dear Dr Rajeev
Thank you for your kind answer
i will soon send you an overview of the work
Best regards
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