| 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 wereincubated 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. To avoid precipitation compound concentration were decreased and the volume of reactionwas high (200 µL). Precipitation probability was less thus addition of detergents was not needed. |
| Affinity data for this assay | |
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