| Field | Specification |
|---|---|
| Target | |
| Alternative names | Osteopontin;Bone sialoprotein 1;Nephropontin;Secreted phosphoprotein 1;SPP-1;Urinary stone protein;Uropontin;SPP1;BNSP, OPN;PSEC0156; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: I17-N300 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Milk, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human osteopontin (gene SPP1) in cell culture supernatants, serum, plasma (heparin, EDTA), human milk and urine. Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
Osteopontin (SPP1) is a secreted, phosphorylated glycoprotein first identified as a major non-collagenous protein of bone, where it binds hydroxyapatite and mediates cell-matrix interactions. It also acts as a cytokine that promotes interferon-γ and IL-12 production, and it is widely studied in bone remodeling, inflammation, fibrosis and tumor progression.
Kit Components
| Anti-Human SPP1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human SPP1 Standard | 2 vials, 10 ng/tube |
| Human SPP1 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA), human milk and urine |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant human SPP1 (Ile17–Asn300), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.6–7.8% (3 samples, n = 16 each) |
| Inter-assay CV | 5.3–8.3% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:100. In the supplier’s tests, plasma samples measured 39–234 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.022; 156 = 0.09; 312 = 0.149; 625 = 0.266; 1250 = 0.524; 2500 = 0.956; 5000 = 1.601; 10000 = 2.089. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 156 | 312 | 625 | 1250 | 2500 | 5000 | 10000 |
| O.D. | 0.022 | 0.09 | 0.149 | 0.266 | 0.524 | 1.056 | 1.901 | 2.389 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 305 | 1771 | 4985 | 314 | 1611 | 5408 |
| Standard deviation | 14.03 | 138.13 | 304.08 | 16.64 | 133.71 | 383.96 |
| CV (%) | 4.6% | 7.8% | 6.1% | 5.3% | 8.3% | 7.1% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
Zhang Junfeng et al. (2025) POU5F1 upregulates SPP1 to sustain stem-like traits and malignant phenotypes via JAK1/STAT3 in clear cell renal cell carcinoma. International Journal of Surgery. 10.1097/JS9.0000000000004469
Xing Qin et al. (2018) Cancer-associated Fibroblast-derived IL-6 Promotes Head and Neck Cancer Progression via the Osteopontin-NF-kappa B Signaling Pathway. Theranostics. 10.7150/thno.22182
Heng Zhang et al. (2026) The CSF1+ tumor cell–SPP1+ macrophage axis drives gastric cancer progression and immunotherapy resistance. Frontiers in Immunology. 10.3389/fimmu.2026.1817573
Xiaomei Yang et al. (2025) Osteopontin regulates right ventricular failure through integrin ανβ3/PERK/CHOP-dependent inflammatory and apoptotic pathways. Frontiers in Immunology. 10.3389/fimmu.2025.1569210
Jiao Mengfan et al. (2026) P-STAT3-dependent SPP1 transcription in cholangiocytes drives hepatic stellate cells activation in congenital hepatic fibrosis. Communications Biology. 10.1038/s42003-026-10476-6
Junyu Cao et al. (2018) Hypoxia-driven paracrine osteopontin/integrin αvβ3 signaling promotes pancreatic cancer cell epithelial–mesenchymal transition and cancer stem cell-like properties by modulating forkhead box protein M1. Molecular Oncology. 10.1002/1878-0261.12399
Yi-Xuan Chen et al. (2019) Diminished membrane recruitment of Akt is instrumental in alcohol-associated osteopenia via the PTEN/Akt/GSK-3β/β-catenin axis. FEBS Journal. 10.1111/febs.14754
Liping Sun et al. (2017) Three-dimensional combined biomarkers assay could improve diagnostic accuracy for gastric cancer. Scientific Reports. 10.1038/s41598-017-12022-1
Baisong Zhao et al. (2020) Combination of nanolamellae and PDA coating on promoting the long-term adhesion, proliferation, and differentiation of osteoblasts. Polymer. 10.1016/j.polymer.2020.122462
Xu Baofeng et al. (2025) Impact of biomarkers on carotid artery disease and stroke. Scientific Reports. 10.1038/s41598-025-13517-y
