Description
Urease Activity Assay Kit (BA0061) (BA0061)
The Urease Activity Assay Kit (SKU: BA0061) provides a very sensitive and convenient means of measuring urease activity in a wide variety of samples, including soil. Urease (amidohydrolase, EC 3.5.1.5) catalyses the hydrolysis of urea into carbon dioxide and ammonia, and many gastrointestinal and urinary tract pathogens produce the enzyme, making its activity a useful diagnostic parameter for pathogens such as Helicobacter pylori. Urease is found in bacteria, yeast and higher plants, and its activity is commonly determined in anaerobes of the bovine rumen, human faeces and environmental samples such as soils and phytoplankton. In this assay urease reacts with urea to form ammonia, which is determined by the Berthelot method at 670 nm. The assay is simple, sensitive, stable and readily adaptable to high-throughput screening.
| Product Name: | Urease Activity Assay Kit (BA0061) |
| SKU: | BA0061 |
| Detection Method: | Colorimetric |
| Detection Range: | As low as 0.003 U/L; up to 25 U/L in the standard assay |
| Sample Type: | Biological and environmental samples including soil |
| Species Reactivity: | All |
| Assay Time: | 10 minute enzyme reaction plus 30 minute detection |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | 4C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Room Temperature |
Quantitative determination of urease activity. Urease reacts with urea to generate ammonia, which is measured by the Berthelot method at 670 nm.
- Safe, non-radioactive assay
- Sensitive: as low as 0.003 U/L urease activity can be quantified
- Homogeneous, convenient mix-incubate-measure format with no wash or transfer steps
- Robust and amenable to high-throughput screening
- Urease activity determination in biological and environmental samples
- Evaluation and screening of urease inhibitors
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Assay preparation. Bring all components to room temperature. For the calibration curve, prepare a 500 uM premix by mixing 5 uL 50 mM NH4Cl with 495 uL Buffer and prepare the dilution series shown in the table. Transfer 90 uL of each standard into separate wells of a clear flat-bottom 96-well plate. Dilute samples in Assay Buffer and transfer 90 uL into separate wells; use 90 uL enzyme buffer as a Sample Blank. |
| 2 | Enzyme reaction. Add 10 uL Urea to each well and incubate at the desired temperature for 10 minutes. |
| 3 | Detection. Add 100 uL Reagent A to each well (this terminates the urease reaction) and tap to mix, then add 50 uL Reagent B and tap to mix again. Incubate for 30 minutes in the dark and read optical intensity at 670 nm (630-700 nm). |
Plot the NH4Cl calibration curve and determine its slope. Urease activity = (ODsample - ODblank) / (slope x t) U/L, where t is the incubation time (10 minutes for the standard assay). One unit of urease catalyses the formation of 1 umole ammonia per minute at pH 7.0 under the assay conditions. If activity exceeds 25 U/L, dilute the enzyme, repeat and multiply by the dilution factor.
| Component | Quantity | Storage |
| Assay Buffer (pH 7.0) | 20 mL | 4C |
| Urea | 1.5 mL | 4C |
| NH4Cl (50 mM) | 100 uL | 4C |
| Reagent A | 12 mL | 4C |
| Reagent B | 6 mL | 4C |
