Human Myeloperoxidase DuoSet ELISA

Catalog # Availability Size / Price Qty
DY3174
Ancillary Products Available
Human Myeloperoxidase / MPO ELISA Standard Curve
1 Image
Product Details
Procedure
Citations (8)
FAQs
Supplemental Products
Reviews (3)

Human Myeloperoxidase DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human Myeloperoxidase. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

 

  

Data Example

Human Myeloperoxidase / MPO ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human Myeloperoxidase DuoSet ELISA

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.

8 Citations: Showing 1 - 8
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  1. Role of NOD1/NOD2 receptors in Fusobacterium nucleatum mediated NETosis
    Authors: HM Alyami, LS Finoti, HS Teixeira, A Aljefri, DF Kinane, MR Benakanake
    Microb. Pathog., 2019;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Calcineurin inhibitor Tacrolimus impairs host immune response against urinary tract infection
    Authors: D Emal, E Rampanelli, N Claessen, FJ Bemelman, JC Leemans, S Florquin, MC Dessing
    Sci Rep, 2019;9(1):106.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Activation of a TLR9 mediated innate immune response in preeclampsia
    Authors: RD Williamson, FP McCarthy, LC Kenny, CM McCarthy
    Sci Rep, 2019;9(1):5920.
    Species: Human
    Sample Types: Plasma
  4. Myeloperoxidase in Hypertensive Disorders of Pregnancy and Its Relation With Nitric Oxide
    Authors: L Rocha-Penh, M Caldeira-D, JE Tanus-Sant, R de Carvalh, VC Sandrim
    Hypertension, 2017;0(0):.
    Species: Human
    Sample Types: Plasma
  5. Possible involvement of NETosis in inflammatory processes in the eye: Evidence from a small cohort of patients
    Authors: T Barliya, R Dardik, Y Nisgav, M Dachbash, D Gaton, G Kenet, R Ehrlich, D Weinberger, T Livnat
    Mol. Vis., 2017;23(0):922-932.
    Species: Human
    Sample Types: Vitreous Humor
  6. Swimming Motility Mediates the Formation of Neutrophil Extracellular Traps Induced by Flagellated Pseudomonas aeruginosa
    PLoS Pathog., 2016;12(11):e1005987.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Therapy with plasma purified alpha1-antitrypsin (Prolastin(R)) induces time-dependent changes in plasma levels of MMP-9 and MPO.
    Authors: Koepke J, Dresel M, Schmid S, Greulich T, Beutel B, Schmeck B, Vogelmeier C, Janciauskiene S, Koczulla A
    PLoS ONE, 2015;10(0):.
    Species: Human
    Sample Types: Serum
  8. Release of cystic fibrosis airway inflammatory markers from Pseudomonas aeruginosa-stimulated human neutrophils involves NADPH oxidase-dependent extracellular DNA trap formation.
    Authors: Yoo D, Winn M, Pang L, Moskowitz S, Malech H, Leto T, Rada B
    J Immunol, 2014;192(10):4728-38.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

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Reviews for Human Myeloperoxidase DuoSet ELISA

Average Rating: 4.3 (Based on 3 Reviews)

5 Star
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4 Star
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3 Star
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Human Myeloperoxidase DuoSet ELISA
By Anonymous on 08/14/2019
Application: Sample Tested: EDTA Plasma

Very pleased with this kit. Very reliable. Used EDTA plasma diluted 1/200 with RD buffer on a 384-well plate format.


Human Myeloperoxidase DuoSet ELISA
By Anonymous on 03/15/2018
Application: Sample Tested: EDTA Plasma

Human Myeloperoxidase DuoSet ELISA
By Anonymous on 06/09/2017
Application: Sample Tested: human serum

Serum samples require a 50-fold diluition