Human Osteopontin ELISA Kit (ab269374)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 88 pg/ml
- Range: 109.38 pg/ml - 7000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Milk, Serum, Urine
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human Osteopontin ELISA Kit
See all Osteopontin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 3 2.1% Inter-assay Sample n Mean SD CV% Serum 8 2.8% -
Sample type
Cell culture supernatant, Milk, Urine, Serum, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
88 pg/ml -
Range
109.38 pg/ml - 7000 pg/ml -
Recovery
Sample specific recovery Sample type Average % Range Cell culture supernatant 108 102% - 115% Milk 96 93% - 98% Urine 98 96% - 100% Serum 102 99% - 104% Hep Plasma 93 90% - 96% EDTA Plasma 99 95% - 102% Cit plasma 92 91% - 93% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human Osteopontin ELISA Kit (ab269374) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Osteopontin protein in edta plasma, hep plasma, milk, serum, urine, cit plasma, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human Osteopontin with 88 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Osteopontin (OPN) is a 35kDa secreted adhesive glycophosphoprotein expressed mainly in bone, teeth, kidney, epithelial lining tissues, activated T cells, macrophages and breast milk. It is processed by matrix metalloproteases 3 and 4 as well as thrombin. It can be present as a cytokine-like soluble protein or as an immobilized extracellular matrix protein. It is important in multiple physiologic and pathologic processes including survival, adhesion, migration, angiogenesis, apoptosis and inflammation. High levels of Osteopontin is used as a biomarker of poor prognosis in patients with breast cancer, multiple myeloma and prostate cancer. Osteopontin has a role in the immune response as a chemotactic factor for macrophages, dendritic cells and T cells. It influences both cell-mediated and humoral immunity and Th1-cytokine functions as well as enhancing B lymphocytes proliferation and immunoglogulin production.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human Osteopontin Capture Antibody 1 x 600µl 10X Human Osteopontin Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent 4BI 1 x 6ml Human Osteopontin Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Binds tightly to hydroxyapatite. Appears to form an integral part of the mineralized matrix. Probably important to cell-matrix interaction.
Acts as a cytokine involved in enhancing production of interferon-gamma and interleukin-12 and reducing production of interleukin-10 and is essential in the pathway that leads to type I immunity. -
Tissue specificity
Bone. Found in plasma. -
Sequence similarities
Belongs to the osteopontin family. -
Post-translational
modificationsExtensively phosphorylated on clustered serine residues.
N- and O-glycosylated.
Phosphorylation sites are present in the extracelllular medium. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- BNSP
- Bone sialoprotein 1
- Bone sialoprotein I
see all -
Database links
- Entrez Gene: 6696 Human
- Omim: 166490 Human
- SwissProt: P10451 Human
- Unigene: 313 Human
Images
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SimpleStep ELISA Protocol Diagram
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Human Osteopontin standard curve comparisonStandard Curve comparison between human Osteopontin SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.
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Example of human Osteopontin standard curve in Sample Diluent NS.The Osteopontin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native Osteopontin in human serum and plasma samples.The concentrations of Osteopontin were measured in duplicates, interpolated from the Osteopontin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 12.5%, plasma (EDTA) 12.5%, and plasma (heparin) 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Osteopontin concentration was determined to be 16.2 ng/mL in serum, 13.5 ng/mL in plasma (citrate), 13.0 ng/mL in plasma (EDTA), and 19.9 ng/mL in plasma (heparin).
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Interpolated concentrations of native Osteopontin in human urine and stimulated PBMC cell culture supernatant samples.The concentrations of Osteopontin were measured in duplicates, interpolated from the Osteopontin standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1:400 and stimulated PBMC cell culture supernatant 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Osteopontin concentration was determined to be 1,516 ng/mL in urine and 4.2 ng/mL in stimulated PBMC cell culture supernatant. No detectable signal was observed in unstimulated PBMC cell culture supernatant.
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Interpolated concentrations of native Osteopontin in human milk samples.The concentrations of Osteopontin were measured in duplicates, interpolated from the Osteopontin standard curves and corrected for sample dilution. Undiluted samples are as follows: milk 1:20,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Osteopontin concentration was determined to be 88 µg/mL in urine.
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Serum from ten individual healthy human female donors was measured in duplicateInterpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Osteopontin concentration was determined to be 19 ng/mL with a range of 10 – 39 ng/mL.
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Sandwich ELISA - Human Osteopontin ELISA Kit (ab269374)To learn more about the advantages of recombinant antibodies see here.
