Human Osteopontin (OPN) ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DOST00
Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Urine (50 µL), Human Milk (50 µL)
Sensitivity
0.024 ng/mL
Assay Range
0.312-20 ng/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Urine, Human Milk)
Product Summary for Human Osteopontin (OPN) Quantikine ELISA Kit
The Quantikine Human Osteopontin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Osteopontin in cell culture supernates, plasma, urine, and human milk. It contains NS0-expressed recombinant human Osteopontin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Osteopontin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Osteopontin.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
Specificity
Natural and recombinant human Osteopontin
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Although serum has been validated for use in this assay, it is not a recommended sample
type because of proteolytic cleavage by thrombin during the clotting process. Serum
values are approximately 50% of the plasma values. See references 32-34 for more
details.
Cell Culture Supernates:
HepG2 human hepatocellular carcinoma cells were cultured in MEM supplemented with 5% fetal bovine serum until confluent. Cells were then stimulated with 50 ng/mL PMA for 24 hours. An aliquot of the cell culture supernate was removed, assayed for human OPN and measured 790 ng/mL.
Plasma/Urine - Samples from apparently healthy volunteers were evaluated for the presence
of human OPN in this assay. No medical histories were available for the donors used in this
study.
| Sample Type | Mean (ng/mL) | Range (ng/mL) | Standard Deviation (ng/mL) |
| EDTA plasma (n=40) | 94.8 | 49.2-175 | 24.9 |
| Heparin plasma (n=40) | 96.2 | 53.4-195 | 26.7 |
| Urine (n=21) | 2044 | 122-8796 | 1951 |
Cell Culture Supernates:
Human peripheral blood cells (1 x 106
cells/mL) were cultured in DMEM supplemented with
5% fetal bovine serum, 5 μM beta-mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with
10 μg/mL PHA. Aliquots of the cell culture supernates were removed and assayed for levels of
human OPN.
| Condition | Day 1 (ng/mL) | Day 6 (ng/mL) |
| Unstimulated | 0.675 | 1.85 |
| Stimulated | 1.50 | 2.67 |
HepG2 human hepatocellular carcinoma cells were cultured in MEM supplemented with 5% fetal bovine serum until confluent. Cells were then stimulated with 50 ng/mL PMA for 24 hours. An aliquot of the cell culture supernate was removed, assayed for human OPN and measured 790 ng/mL.
A431 human epithelial carcinoma cells were cultured in DMEM supplemented with
10% 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 OPN and
measured 8.54 ng/mL.
Human Milk - Three human milk samples were assayed for human OPN and ranged from
35,600-248,000 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, EDTA Plasma, Heparin Plasma, Human Milk, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (ng/mL) | 2.30 | 4.90 | 9.30 | 2.36 | 4.81 | 9.17 |
| Standard Deviation | 0.092 | 0.126 | 0.267 | 0.155 | 0.274 | 0.499 |
| CV% | 4.0 | 2.6 | 2.9 | 6.6 | 5.7 | 5.4 |
Recovery for Human Osteopontin (OPN) Quantikine ELISA Kit
The recovery of Osteopontin spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 104 | 86-113 |
| EDTA Plasma (n=4) | 102 | 92-114 |
| Heparin Plasma (n=4) | 101 | 95-112 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Osteopontin were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Osteopontin (OPN) Quantikine ELISA Kit
Human Osteopontin/OPN ELISA Standard Curve
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: Osteopontin/OPN
Long Name
Secreted Phosphoprotein 1 [BNSP]
Alternate Names
Eta-1, OPN, Spp1
Gene Symbol
SPP1
Additional Osteopontin/OPN Products
Product Documents for Human Osteopontin (OPN) Quantikine ELISA Kit
Product Specific Notices for Human Osteopontin (OPN) Quantikine ELISA Kit
For research use only
Loading...
Loading...
Loading...
⚠ 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.