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Human Myeloperoxidase ELISA Kit - Quantikine Best Seller

R&D Systems, part of Bio-Techne | Catalog # DMYE00B

R&D Systems, part of Bio-Techne
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DMYE00B

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Cell Lysates (50 µL), Serum (10 µL), Platelet-poor EDTA Plasma (10 µL), Platelet-poor Heparin Plasma (10 µL), Saliva (10 µL), Urine (50 µL)

Sensitivity

0.062 ng/mL

Assay Range

0.156-10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Saliva, Urine)

Product Summary for Human Myeloperoxidase Quantikine ELISA Kit

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.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human

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.

Sample Values

Serum/Plasma/Saliva/Urine - Samples from apparently healthy volunteers were evaluated for the presence of human MPO in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Serum (n=29)10021.4-22965
Platelet-poor EDTA plasma (n=29)7.822.89-27.44.93
Platelet-poor heparin plasma (n=29)9.473.57-28.45.27
Saliva (n=10)55831.2-1428461
Urine (n=13)4.390.17-13.54.68

Cell Culture Supernates:
Human monocytes were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 1 μg/mL LPS for 1 and 6 days. Aliquots of the cell culture supernates were removed and assayed for human MPO.

ConditionDay 1 (ng/mL)Day 6 (ng/mL)
Unstimulated28.041.4
Stimulated29.930.3

HL-60 human acute promyelocytic leukemia cells were cultured in RPMI supplemented with 20% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human MPO, and measured 18.2 ng/mL.

An aliquot of the same HL-60 cells was removed, and the cells were lysed at 5 x 106 cells/mL in diluted Cell Lysis Buffer 3. The lysate was assayed for human MPO and measured 2673 ng/mL.


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.

Cell Culture Supernates, Cell Lysates, Saliva

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 0.994 3.36 6.68 0.974 2.99 6.23
Standard Deviation 0.034 0.085 0.123 0.078 0.18 0.38
CV% 3.4 2.5 1.8 8.0 6.0 6.0

Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Serum, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 1.03 3.13 6.63 1.07 3.29 6.71
Standard Deviation 0.024 0.046 0.172 0.12 0.26 0.55
CV% 2.3 1.5 2.6 10.8 8.0 8.2

Recovery for Human Myeloperoxidase Quantikine ELISA Kit

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/MPO ELISA Linearity

Scientific Data Images for Human Myeloperoxidase Quantikine ELISA Kit

Human Myeloperoxidase/MPO ELISA Cell Culture Supernate/Cell Lysate/Saliva Standard Curve

Human Myeloperoxidase/MPO ELISA Cell Culture Supernate/Cell Lysate/Saliva Standard Curve

Human Myeloperoxidase/MPO ELISA Serum/Plasma/Urine Standard Curve

Human Myeloperoxidase/MPO ELISA Serum/Plasma/Urine Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. 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.

Alternate Names

MPO

Entrez Gene IDs

4353 (Human); 17523 (Mouse); 303413 (Rat)

Gene Symbol

MPO

Additional Myeloperoxidase/MPO Products

Product Documents for Human Myeloperoxidase Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Myeloperoxidase Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.