| Assay Method Information | |
| | Urease Inhibition Assay |
| Description: | This assay was modified from Berthelot assay and was employed for the determination of urease activity. The assay is based on the hydrolysis of urea into ammonia which reacts with phenol-hypochlorite to form light blue colored complex measured at 625 nm. A total volume of 85 µl assay mixture contained 10 µl of 50 mM phosphate buffer, pH 7.0, 10 µl of sample solution and 25 µl of Bacillus pasteurii urease (Sigma) solution (0.015 units). The contents were pre-incubated at 37 °C for 10 min. Then, 40 µl of urea stock solution (20 mM) was added to each well and incubation continued at 37 °C for further 10 min and preread at 625 nm using the 96-well plate reader Synergy HT (Biotek Inc.). Phenol hypochlorite (115 µl) reagent was added in each well (freshly prepared by mixing 45 µl phenol reagent with 70 µl of alkali reagent). For color development, incubation was done at 37 °C for another 10 min. Absorbance was again measured at 625 nm. |
| Affinity data for this assay | |
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