Alpha Glucosidase Inhibition Assay
Alpha-glucosidase stands as a target for the treatment of several diseases. It is a key enzyme in carbohydrate metabolism. However, it is currently being investigated against other diseases such as diabetes, cancer, hepatitis B, and HIV. Inhibiting alpha glucosidase temporarily delays the intestinal absorption of carbohydrates and therefore suppresses the elevation of postprandial blood glucose levels. Thus, representing a convenient method for the control of type 2 diabetes. Up till now, three approved alpha-glucosidase inhibitors are present in the market i.e., Acarbose, Miglitol and Voglibose. Therefore, research efforts seeking novel inhibitors with improved efficacy and lower side effects are increasing.
The assay is carried out using alpha-glucosidase from Saccharomyces cervisaceae and ortho-nitrophenol glucoside (ONPG) as substrate. Upon hydrolysis by the enzyme, ONPG turns to yellow-colored ortho-nitrophenol which can be measured colorimetrically and correlated to the enzyme activity.
1- Initial screening at two concentration levels (100 and 10 ug/mL or uM)
2- IC50 (5 concentrations).
N.B. kinetic measurement is not available right now