| Assay Method Information | |
| | β-Glucuronidase Inhibition Assay |
| Description: | The protocol followed in this study was earlier reported [33,34]. β-Glucuronidase activity was evaluated by measuring the absorbance at 405 nm of p-nitrophenol formed from the substrate by the spectrophotometeric method. The total reaction volume was set at 250 µL. The reaction mixture comprises of 185 µL of 0.1 M acetate buffer, 5 µL of test compound solution, 10 µL of enzyme solution and incubated at 37 °C for 30 min. The readings were recorded on a multiplate reader (SpectraMax plus 384) at 405 nm after the addition of 50 µL of 0.4 mM p-nitrophenyl-b-Dglucuronide. All assays were carried out in triplicate. Beside this, to prevent precipitation, the concentration of the compounds was decreased and the reaction volume was high (200 µL) so the probability of precipitation was attenuated hence the addition of detergents was not required. |
| Affinity data for this assay | |
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