Myeloperoxidase (MPO) Colorimetric Assay Kit

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

      Catalog number:E-BC-K074-S

      Size:
      • 100 Assays
      Qty:
      - +
      Price: $180

      Detection method: Colorimetric method

      Detection instrument: Spectrophotometer

      Valid period: 6 months

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

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      General information

      Detection significance

      Myeloperoxidase is a heme-containing cationic glycoprotein that belongs to the heme peroxidase family in mammals. MPO is a dimer formed by polymerization of two subunits. Each subunit contains a heavy chain and a light chain. MPO is abundant in the azuropathic granules of polymorphonuclear leukocytes (PMNLs) and a small number in monocytes and macrophages. Studies have shown that MPO plays an important role in the generation of oxidants and host defense in neutrophils and is closely related to the pathogenesis of many diseases, including cardiovascular disease, lung injury and cancer.

      Detection principle

      Myeloperoxidase reduces hydrogen peroxide to a complex. The complex react with o-dianisidine (as hydrogen donor) to produce a yellow product which has a maximum absorption peak at 460 nm. The activity of MPO can be calculated indirectly by measuring the OD value at 460nm.

      The key point

      The supernatant after centrifugation must be clarified.

      Operation procedures

      Operation steps

      1.   Sample pretreatment

      1)    For serum (plasma) and milk sample:

      Take 0.45 mL of sample and add 0.45 of Reagent 2 application solution, mix fully, then add 0.1 of Reagent 3 application solution and incubate at 37℃ for 15 min.

      2)    For tissue an cells sample:

      Take 0.9 mL of sample and add 0.1 of Reagent 3 application solution, mix fully and incubate at 37 for 15 min.

        

      2.   The measurement of samples

      1)    Control tube: Add 3 mL of double distilled water, 0.2 mL of sample, 0.2 mL of Reagent 5 into 5 mL EP tubes.

      Sample tube: Add 0.2 mL of sample, 0.2 mL of Reagent 5, 3 mL of chromogenic agent into 5 mL EP tubes.

      2)    Oscillate fully with a vortex mixer and incubate for 30 min at 37℃.

      3)    Add 0.05 mL of Reagent 8, oscillate fully with a vortex mixer and incubate for 10 min at 60℃.

      4)    Centrifuge the tubes at 2325 × g for 10 min and take the supernatant for measuring the OD value.

      5)    Set the spectrophotometer to zero with double distilled water and measure the OD value of each tube at 460 nm with 1.0 cm optical path cuvette immediately. 

      Operation table

       

      Control tube

      Sample tube

      Double distilled water (mL)

      3

       

      Sample (mL)

      0.2

      0.2

      Reagent 5 (mL)

      0.2

      0.2

      Chromogenic agent (mL)

       

      3

      Mix thoroughly, incubate at 37 water bath for 30 min.

      Reagent 8 (mL)

      0.05

      0.05

      Mix thoroughly and incubate at 60 water bath for 10 min. Centrifuge the tubes at 2325 × g for 10 min and take the supernatant. Set the spectrophotometer to zero with double distilled water and measure the OD value of each tube at 460 nm with 1.0 cm optical path cuvette immediately.

      Performance characteristics

      Technical parameter

      Detection range 4.9-196.7 U/L Average inter-assay CV 9.8%
      Sensitivity 4.9 U/L Average intra-assay CV 4.5%
      Average recovery rate 104%

      Citations

      1. Publication: Yang Y, Yin B, Lv L, et al. Gastroprotective Effect Of Aucubin Against Ethanol-induced Gastric Mucosal Injury In Mice[J]. Life Sciences, 2017, 189: 44-51.
      2. Publication: Yang Y, Wang Z Y, Zhang L, et al. Protective Effect Of Gentiopicroside From Gentiana Macrophylla Pall. In Ethanol-Induced Gastric Mucosal Injury In Mice[J]. Phytotherapy Research, 2017.
        Species: Human
        Sample Type: Tissue homogenate
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