| Assay Method Information | |
| | Urease Inhibition Assay |
| Description: | Reaction mixture consisting of 25 無 of Jack bean (Canavalia ensiformis) urease (1 unit/well), 55 無 of 100 mM urea dissolved in phosphate buffer (4 mM concentration of 6.80 pH, and 5 無 of various concentrations of test compound (from 0.5 to 0.00625 mM) were incubated at 30 蚓 for 15 min in 96-well plates.In kinetic experiments, various concentrations of both substrate and test compounds were used. Subsequently 45 無 phenol reagents (1% w/v phenol and 0.005% w/v sodium nitroprusside), and 70 無 of alkali reagent (0.5% w/v NaOH and 0.1% w/v NaOCl) were added to each well. Urease activity was measured through Weatherburn indophenols method by the production of ammonia [21]. After 50 min duration, the increasing absorbance at 630 nm was measured in a microplate reader (SpectraMax M2, Molecular Devices, CA, USA). All reactions were performed in triplicate in a final volume of 200 lL. Thiourea was used as the standard inhibitor of urease [15]. |
| Affinity data for this assay | |
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