Human Myeloperoxidase ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DMYE00B
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.
Cell Culture Supernates:
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.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| Serum (n=29) | 100 | 21.4-229 | 65 |
| Platelet-poor EDTA plasma (n=29) | 7.82 | 2.89-27.4 | 4.93 |
| Platelet-poor heparin plasma (n=29) | 9.47 | 3.57-28.4 | 5.27 |
| Saliva (n=10) | 558 | 31.2-1428 | 461 |
| Urine (n=13) | 4.39 | 0.17-13.5 | 4.68 |
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.
| Condition | Day 1 (ng/mL) | Day 6 (ng/mL) |
| Unstimulated | 28.0 | 41.4 |
| Stimulated | 29.9 | 30.3 |
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.
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 Serum/Plasma/Urine Standard Curve
Preparation and Storage
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 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.
Additional Myeloperoxidase/MPO Products
Product Documents for Human Myeloperoxidase Quantikine ELISA Kit
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.