Human Myeloperoxidase Quantikine ELISA Kit

(10 citations)   
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), Platelet-poor EDTA Plasma (10 uL), Platelet-poor Heparin Plasma (10 uL), Saliva (10 uL), Urine (50 uL)
  • Sensitivity
    0.062 ng/mL
  • Assay Range
    0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Urine)
  • Specificity
    Natural human MPO
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Human MPO Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human MPO in cell culture supernates, cell lysates, serum, platelet-poor plasma, saliva, and urine.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma, Human Milk
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean15.732.464.11938.369.2
Standard Deviation1.22.54.31.83.24.5
CV%7.57.76.69.48.36.5

Cell Culture Supernates, Cell Lysates, Saliva, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean10.320.439.410.321.742.8
Standard Deviation0.751.53.611.94
CV%7.27.69.1108.89.2

Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean1.033.136.631.073.296.71
Standard Deviation0.0240.0460.1720.120.260.55
CV%2.31.52.610.888.2

Cell Culture Supernates, Cell Lysates, Saliva
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean0.9943.366.680.9742.996.23
Standard Deviation0.0340.0850.1230.0780.180.38
CV%3.42.51.8866

Recovery

The recovery of MPO spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 92-105
Platelet-poor EDTA Plasma (n=4) 95 90-101
Platelet-poor Heparin Plasma (n=4) 91 83-100
Serum (n=4) 100 88-110
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of MPO were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Myeloperoxidase Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Myeloperoxidase/MPO
Myeloperoxidase (MPO) is a heme-containing enzyme belonging to the XPO subfamily of peroxidases. It is an abundant neutrophil and monocyte glycoprotein that catalyzes the hydrogen peroxide dependent formation of hypochlorus acid (HOCl) and other reactive species. Enzymatically active MPO is a disulfide-linked tetramer that contains two heme groups and two copies each of the heavy and light chains. MPO binds Albumin, MMR, Cytokeratin 1 on vascular endothelial cells, HMW Kininogen, and Integrin CD11b/CD18 on neutrophils. These interactions promote MPO clearance, a reduction of nitric oxide and bradykinin levels, reduced vasodilation, and continued neutrophil activation. Neutrophil MPO is stored in cytoplasmic azurophilic granules. Upon cellular activation and degranulation, MPO is delivered into phagosomes where it is required for the killing of phagocytosed bacteria. Activated neutrophils also release granule contents extracellularly. Elevated plasma MPO levels have been associated with a variety of clinical conditions including systemic inflammation, eclampsia, risk of cardiovascular events, vascular endothelial dysfunction, severity of multiple sclerosis, and prospective mortality and oxidative stress during hemodialysis.
    • Entrez Gene IDs
      4353 (Human); 17523 (Mouse); 303413 (Rat);
    • Alternate Names
      EC 1.11.1; EC 1.11.1.7; Lactoperoxidase; MPO; Myeloperoxidase;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. Add Assay Diluent
    4.   For Serum, Plasma, & Human Milk Samples: Add 50 µL of Assay Diluent to each well.
      For Cell Culture Supernate, Cell Lysate, Saliva, & Urine Samples: Add 100 µL Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well.
    10.   For Serum, Plasma, & Human Milk Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
      For Cell Culture Supernate, Cell Lysate, Saliva, & Urine Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour on the shaker.
    11.   Aspirate and wash 4 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes on the benchtop. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
    Citations:

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    10 Citations: Showing 1 - 10
    Filter your results:

    Species
    Sample Type
    1. Complex regulation of neutrophil-derived MMP-9 secretion in central nervous system tuberculosis
      Authors: CW Ong, PJ Pabisiak, S Brilha, P Singh, F Roncaroli, PT Elkington, JS Friedland
      J Neuroinflammation, 2017;14(1):31.
      Species: Human
      Sample Type: Whole Cells
    2. Inflammatory Differences in Plaque Erosion and Rupture in Patients With ST-Segment Elevation Myocardial Infarction
      Authors: S Chandran, J Watkins, A Abdul-Aziz, M Shafat, PA Calvert, KM Bowles, MD Flather, SA Rushworth, AD Ryding
      J Am Heart Assoc, 2017;6(5):.
      Species: Human
      Sample Type: Plasma
    3. The interdependencies of viral load, the innate immune response, and clinical outcome in children presenting to the emergency department with respiratory syncytial virus-associated bronchiolitis
      Authors: FA Piedra, M Mei, V Avadhanula, R Mehta, L Aideyan, RP Garofalo, PA Piedra
      PLoS ONE, 2017;12(3):e0172953.
      Species: Human
      Sample Type: Nasal Lavage Fluid
    4. Combined administration of anisodamine and neostigmine rescued acute lethal crush syndrome through ?7nAChR-dependent JAK2-STAT3 signaling
      Sci Rep, 2016;6(0):37709.
      Species: Mouse
      Sample Type: Serum
    5. Growth differentiation factor-15 (GDF-15) levels are associated with cardiac and renal injury in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass.
      Authors: Kahli A, Guenancia C, Zeller M, Grosjean S, Stamboul K, Rochette L, Girard C, Vergely C
      PLoS ONE, 2014;9(8):e105759.
      Species: Human
      Sample Type: Plasma
    6. Increased levels of leukocyte-derived MMP-9 in patients with stable angina pectoris.
      Authors: Jonasson S, Lundberg A, Kalvegren H, Bergstrom I, Szymanowski A, Jonasson L
      PLoS ONE, 2011;6(4):e19340.
      Species: Human
      Sample Type: Serum
    7. Presence of IL-5 protein and IgE antibodies to staphylococcal enterotoxins in nasal polyps is associated with comorbid asthma.
      Authors: Bachert C, Zhang N, Holtappels G, De Lobel L, Van Cauwenberge P, Liu S, Lin P, Bousquet J, Van Steen K
      J. Allergy Clin. Immunol., 2010;126(5):962.
      Species: Human
      Sample Type: Tissue Homogenates
    8. Effects of atorvastatin added to inhaled corticosteroids on lung function and sputum cell counts in atopic asthma.
      Authors: Hothersall EJ, Chaudhuri R, McSharry C, Donnelly I, Lafferty J, McMahon AD, Weir CJ, Meiklejohn J, Sattar N, McInnes I, Wood S, Thomson NC
      Thorax, 2008;63(12):1070-5.
      Species: Human
      Sample Type: Sputum
    9. Interleukin-8 is increased in the membrane of circulating erythrocytes in patients with acute coronary syndrome.
      Authors: Tziakas DN, Chalikias GK, Tentes IK, Stakos D, Chatzikyriakou SV, Mitrousi K, Kortsaris AX, Kaski JC, Boudoulas H
      Eur. Heart J., 2008;29(22):2713-22.
      Species: Human
      Sample Type: Plasma
    10. Computed tomographic scan-diagnosed chronic obstructive pulmonary disease-emphysema: eotaxin-1 is associated with bronchodilator response and extent of emphysema.
      Authors: Miller M, Ramsdell J, Friedman PJ, Cho JY, Renvall M, Broide DH
      J. Allergy Clin. Immunol., 2007;120(5):1118-25.
      Species: Human
      Sample Type: BALF
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