| Assay Method Information | |
| | Urease Inhibition Assay |
| Description: | The urease inhibitory activity of synthesized sulfonamides (3a-j) was determined by measuring the amount of ammonia produced by the indophenols method described byWeatherburn (27,28). The reaction mixtures, comprising 20 µL of enzyme (Jack bean urease, 5 U/mL) and 20 µL of test compounds in 50 µL buffer (100 mM urea, 0.01 MK2HPO4, 1 mM EDTA and 0.01 M LiCl2, pH 8.2), were incubated for 10 min at 37 °C in 96-well plate. Briefly, 40 µL each of phenol reagents (1%, w/v phenol and0.005%, w/v sodium nitroprusside) and 70 µL of alkali reagent (0.5%, w/v NaOH and 0.1% active chloride NaOCl) were added to each well. The absorbance at 625 nm was measured after 30 min, using a microplate reader (OPTIMax, Tunable). All reactions were performed in triplicate. |
| Affinity data for this assay | |
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