Assay Method Information

Assay Name:  β-Glucuronidase Inhibition Assay
Description:  β-Glucuronidase inhibitory activity was measured by quantifying the absorbance of para-nitrophenol which was formed from the substrate by the spectrophotometricmethod at 405 nm. Total volume of reaction was 250 μL. DMSO (100%) was used to solublized the compound (5 μL) which become 2% in the final assay (250 μL)and the same conditions were used for standard (D-saccharic acid 1,4-lactone). Reaction mixture comprised of 0.1 M acetate buffer (185 μL), test compound solution in DMSO (5 μL), enzyme solution (10 μL), was incubated for 30 min at 37 °C. Than 0.4 mM pnitrophenyl-β-D-glucuronide (50 μL) was added and plate wasread on a multiplate reader (SpectraMax plus 384, USA) at 405 nm. All assays were run in triplicate.
Affinity data for this assay
 

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