| Assay Method Information | |
| | Urease Inhibition Assay |
| Description: | Reaction mixtures comprising 25 μL of enzyme (jack bean urease) solution and 55 μL of buffers containing 100 mM urea were incubated with 5 μL of test compounds (0.5 mM concentration) at 30°C for 15 min in 96-well plates. Urease activity was determined by measuring ammonia production using the indophenol method as described by weather burn14. Briefly, 45 μL each phenol reagent (1% w/v phenol and 0.005% w/v sodium nitroprusside) and 70 μL of alkali reagent (0.5% w/v NaOH and0.1% active chloride NaOCl) were added to each well. The increasing absorbance at 630 nm was measured after 50 min, using a microplate reader (Molecular Device). All reactions were performed in triplicate in a final volume of 200 μL. |
| Affinity data for this assay | |
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