I had tested Acarbose in Alpha-glucosidase assay, the result showed a very low absorbance, it almost same reading as negative control (Distilled water). Below is my method of the assay. Anyone knows what is the solution to solve it?
1. 650 ul buffer + 100 ul acarbose (0.2-1mg/ml)
2. add 100 ul enzyme( 0.5 U/mL)
3. incubate at 37C for 15 min
4. add 350 ul pNPG (10 mM)
5. incubate at 37C for 30 min
6. add 250 ul of Na2CO3 (0.1 M)
7. read at 405 nm
If you need to measure alpha-glucosidase activity don't add Acarbose because it's an ihibitor of glucosidases.
If your purpose is to measure the inhibitory effect on your enzyme. First, be sure that you enzyme is active. After that, use different concentrations of the inhibitor but with kinetic measurement of the activity.
Get the basic informations about Acarbose.
Hi Noureddine Boulenouar, i want to measure the inhibitory effect of enzyme. Yup, the enzyme is active. Could you explain about the kinetic measurement of the activity?
I found that Acarbose got different brands, will it affect the result using different brand of acarbose? Thank you.
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