| Assay Method Information | |
| | β-Glucuronidase Assay |
| Description: | β-glucuronidase activity was determined by measuring absorbance at 405 nm of p-nitrophenol formed substrate by spectrophotometric method. 250 μL was the volume of total reaction. Reaction mixture containing 5 μL of test compound solution, 185 μL of 0.1 M acetate buffer and 10 μL of enzyme solution were incubated for 30 min at 37 °C. At 405 nm the plates were recorded on multiplate reader (SpectaMax plus 384) after the addition of 50 μL of 0.4 mM p-nitrophenyl-β-D-Glucuronide. Experiments were performed for triplicate [Khan et al., Bioorg. Med. Chem., 22:3449; Khan et al., J. Comput. Aided Mol. Des., 28:577-585]. To avoid precipitation compound concentration were decreased and the volume of reaction was high (200 μL). Precipitation probability was less thus addition of detergents was not needed. |
| Affinity data for this assay | |
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