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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL), Human Milk (50 uL)
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Sensitivity
0.024 ng/mL
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Assay Range
0.3 - 20 ng/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Urine, Human Milk)
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Specificity
Natural and recombinant human Osteopontin
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC136, Quantikine Immunoassay Control Set 960 for Human Osteopontin -
Product Summary
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.
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, Urine, Human Milk | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 2.3 | 4.9 | 9.3 | 2.36 | 4.81 | 9.17 |
| Standard Deviation | 0.092 | 0.126 | 0.267 | 0.155 | 0.274 | 0.499 |
| CV% | 4 | 2.6 | 2.9 | 6.6 | 5.7 | 5.4 |
Recovery
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.
Preparation and Storage
Background: Osteopontin/OPN
Osteopontin (OPN), also known as bone sialoprotein (BSP), is a secreted SIBLING family protein that can be variably modified by O- and N-glycosylation, sulfation, phosphorylation, and transglutamination. OPN is widely expressed and is prominent in mineralized tissues. It inhibits bone mineralization and kidney stone formation and promotes inflammation, cell ad¬hesion, and migration. Its expression is upregulat¬ed during inflammation, obesity, atherosclerosis, cancer, and tissue damage and contributes to the pathophysiology of these conditions. The central region of OPN contains RGD and non-RGD binding sites for multiple integrins. Adjacent to the RGD motif is the sequence SLAYGLR (SVVYGLR in human) which serves as a cryptic binding site for additional integrins: it is masked in full length OPN but is exposed following OPN cleavage by multiple proteases in tumors and sites of tissue injury.
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Alternate Names
BNSP; Bone sialoprotein 1; Eta-1; MGC110940; Nephropontin; OPN; Osteopontin; secreted phosphoprotein 1bone sialoprotein I, early T-lymphocyteactivation 1); secreted phosphoprotein-1 (osteopontin, bone sialoprotein); Spp1; SPP-1; SPP1/CALPHA1 fusion; Urinary stone protein; uropontin;
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