Urea Colorimetric Assay Kit (Urease method)

    • Biochemical Assay Kit-Elabscience
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    • Biochemical Assay Kit-Elabscience
    • Biochemical Assay Kit-Elabscience

      Catalog number:E-BC-K183

      Size:
      • 100 Assays
      • 50 Assays
      Qty:
      - +
      Price: $150

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometry (Visible range)

      Valid period: 3 months

      Lead Time: 7~10 daysWelcome to order from local distributors.

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      Application

      This kit can be used to measure urea content in animal serum, plasma, urine, saliva, milk and other samples. 

       

      Detection significance

      Urea is the major final-product of protein metabolism in the body, which constitutes the clear majority of non-protein nitrogen in blood. Blood urea nitrogen come from the liver, which excreted with urine through kidney. Renal function failure, nephritis, urinary tract obstruction and so on can make the content of blood urea increased.

       

      Detection principle

      Urea can be decomposed into ammonia ion and carbon dioxide by urease. Ammonia ion can react with amphyl and form a green substance in alkaline medium, and the production of the blue substance is proportional to the urea content which can be calculated with the colorimetric assay at 580 nm.


      Experimental instruments

      Test tube, Micropipettor, Vortex mixer, 37water bath, Centrifuge, Spectrophotometer (580 nm)


      Preparation of Sample

      1. Plasma (serum): Detect directly. Samples can be store at room temperature for 24 hours or at 4 for 7 days. Use oxalate, heparin or EDTA as the plasma anticoagulant.

      2. Urine: Collect the fresh urine and centrifuge the sample at 10000 g for 10 min at 4℃. Take the supernatant and dilute the urine with normal saline at a ratio of 1:19~1:50. Increase the dilution multiple of sample if the result exceeds the linear range.

      3. Saliva: Gargle with clear water, collect the saliva 30 min later, centrifuge at 10000 g for 10 min at 4. Take the supernatant and preserve it on ice for detection.

      4. Milk: Collect fresh milk, centrifuge at 10000 g for 10 min at 4℃, Remove the upper layer of milky white, take the middle layer supernatant and preserve it on ice for detection

       

      Operation steps

      (1) Blank tube: Add 0.02 mL of Double-distilled water into a 5 mL centrifuge tube.

      Standard tube: Add 0.02 mL of 10 mmol/L urea standard working solution into a 5 mL centrifuge tube.

      Sample tube: Add 0.02 mL of Sample into a 5 mL centrifuge tube.

      Control tube: Add 0.02 mL of Sample into a 5 mL centrifuge tube.

      (2) Add 0.25 mL of enzyme working solution to blank tube, standard tube and sample tube, add 0.25 mL of reagent 3 to blank tube, mix fully with vortex mixer.

      (3) Add 1 mL of reagent 4 and 1 mL of reagent 5, mix fully, incubate at 37water bath for 10 min.

      (4) Set to zero with double distilled water and measure the OD values of each tube with 1 cm cuvette diameter at 580 nm.

       

      Note: The following operating table could be as a reference.


       

      Blank tube

      Standard tube

      Sample tube

      Control tube

      Double-distilled water (mL)

      0.02

       

       

       

      10 mmol/L urea standard working solution (mL)

       

      0.02

       

       

      Sample (mL)

       

       

      0.02

      0.22

      Enzyme working solution (mL)

      0.25

      0.25

      0.25

       

      Reagent 3 (mL)

       

       

       

      0.25

      Mix fully, incubate in 37 water bath for 10 min accurately.

      Reagent 4 (mL)

      1

      1

      1

       

      Reagent 5 (mL)

      1

      1

      1

       

      Mix fully and incubate in 37 water bath for 10 min. Set to zero with double distilled water and measure the OD values of each tube with 1 cm cuvette diameter at 580 nm.

      Technical parameters

      1. The sensitivity of the kit is 0.114 mmol/L.

      2. The intra-assay CV is 4.6% and the inter-assay CV is 4.7%.

      3. The recovery of the kit is 102%.

      4. The linear range of the kit is 0.114-30 mmol/L.

       

      Note

      1. The kit is for scientific research only.

      2. Instructions should be followed strictly, changes of operation may result in unreliable results.

      3. The validity of kit is 3 months.

      4. Do not use components from different batches of kit.

      5. Properly dilute the sample if the color is too dark, and multiply by dilution factor when calculating the result.

      6. It is recommended to use disposable plastic tubes to avoid contamination.

      7. Prepare fresh enzyme working solution for needed amount before use. The enzyme working solution cannot be store for a long time.

      8. The adhesion of enzyme stock solution is strong. It should be slowly absorbed when absorbing with pipette.

      9. The incubation time must be 10 min accurately after adding enzyme working solution. Therefore, it is better to make batch operation if there are many samples to be detected. The number of operation in a batch should be less than 20.


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