Urease Assay Kit

Produktnummer ABIN1000280

Produktdetails

Target: Urease (URE)

Applikation: Activity Assay (AcA)

Spezifität: 0.0014 U/L

Produktmerkmale: Safe. Non-radioactive assay.
Sensitive and accurate. As low as 0.003 U/L urease activity can be quantified.
Homogeneous and convenient. Mix-incubate-measure type assay. No wash and reagent transfer steps are involved.
Robust and amenable to HTS: can be readily automated on HTS liquid handling systems for processing thousands of samples per day.

Bestandteile: Assay Buffer: 20 mL (pH 7.0). Reagent A: 12 mL. Urea: 1.5 mL. Reagent B: 6 mL. NH4Cl: 100 µL 50 mM.

Benötigtes Material: Pipetting devices, centrifuge tubes, clear flat-bottom 96-well plate (e.g. Corning Costar).

Anwendungsinformationen

Applikationshinweise: Urease activity determination in biological and environmental samples. Evaluation and screening for urease inhibitors.

Kommentare:

(1). Soil and other environmental samples can be extracted in Assay Buffer (10 mM sodium phosphate, pH 7.0) using any established methods. For such low urease activity samples, incubate the urease reaction for 2 to 4 hours at 30 or 37 °C (Step 2). Soil samples may contain very low concentrations of ammonia. To correct for sample ammonia, immediately prior to detection (Step 3), prepare Sample Blank by mixing the following in this order: 100 μL Reagent A, 90 μL sample extract, 10 μL urea and 50 μL reagent B.
(2). Cuvet assays: scale up 4-fold to a total of 1 mL reaction by using 360 μL Sample, 40 μL Urea, 400 μL Reagent A and 200 μL Reagent B.

Protokoll: Interference: ammonia is known to interfere with this assay and prior to assay, should be removed by dialysis or filtration.
1. Assay Preparation. Prior to assay, bring all components to room temperature. For calibration curve, prepare a 500 µMpremix by mixing 5 µL 50mM NH4Cl and 495 µL Buffer. Dilute NH4Cl. Transfer 90 µL into separate wells of a clear flat-bottom 96-well plate. Samples. Dilute sample in Assay Buffer (Note: it is prudent to test different dilutions to ensure urease activity is within the detection range). Transfer 90 µL sample into separate wells. Use 90 µL enzyme buffer as a Sample Blank.
2. Enzyme Reaction. Add 10 µL Urea to each well. Incubate at desired temperature for 10 min.
3. Detection. Add 100 µL Reagent A to each well. Tap plate to mix. Then add 50 µL Reagent B to each well. Tap plate to mix again. Note: addition of Reagent A terminates the urease reaction. Incubate for 30 min in the dark. Read optical intensity at 670nm (630- 700nm).

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: 4 °C

Antigendetails

Target: Urease (URE)

Andere Bezeichnung: Urease (URE Produkte)

Hintergrund: Quantitative determination of urease activity by colorimetric (630nm) method.
Procedure: 40 min.

Urease (Amidohydrolase, EC 3.5.1.5) is an enzyme that catalyzes the hydrolysis of urea into carbon dioxide and ammonia. Many gastrointestinal or urinary tract pathogens produce urease. Thus its activity is a useful diagnostic parameter for the presence of pathogens such as Helicobacter pylori. Urease is found in bacteria, yeast, and higher plants. Urease activity is commonly determined in anaerobes of the bovine rumen, human feces and environmental samples such as soils and phytoplanktons. This Urease Assay Kit provides a very sensitive and convenient means to measure urease activity in a variety of samples including soil. In the assay, urease reacts with urea, resulting in the formation of ammonia, which is determined by the Berthelot method at 670nm. The assay is simple, sensitive, stable and high-throughput adaptable.