I am planning to assay the effect of plant extracts on the activity of anti-glucosidase in vitro
Dear Researcher
Here is a brief method for deremination of glucosidase inhibition
Alpha glucosidase (2U/ml) was premixed with 20 μl of test sample at various concentrations (10, 50 and 100 μg/ml) and incubated for 5 min at 37ºC. 1mM para-nitrophenyl glucopyanoside (20 μl) in 50mM of phosphate buffer (pH 6.8) was added to initiate the reaction. The mixture was further incubated at 37ºC for 20 min. The reaction was terminated by addition of 50 μl of 1 M sodium carbonate and the final volume was made up to 150 μl. Alpha glucosidase activity was determined spectrophotometrically at 405nm by measuring the quantity of para-nitrophenol released from pNPG. The assay was performed in triplicate. The concentration of test sample required to inhibit 50% of alpha glucosidase activity under the conditions was defined as the IC50 value.
Method is adopted from: Pistia-Brueggeman G., Hollingsworth R.I. (2007). A preparation and screening strategy for glycosidase inhibitors. Tetrahedron, 57:8773-8778
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