| Field | Specification |
|---|---|
| Target | |
| Alternative names | Putative uncharacterized protein ;Spp1 ; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: L17-N294 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma, Urine |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies mouse osteopontin (gene Spp1) in cell culture supernatants, serum, plasma (heparin, EDTA) 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-Mouse Spp1 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Spp1 Standard | 2 vials, 10 ng/tube |
| Mouse 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 | Mouse |
|---|---|
| Sample types | Cell culture supernatants, serum, plasma (heparin, EDTA) and urine |
| Assay range | 156 pg/mL–10,000 pg/mL |
| Sensitivity | <10 pg/mL |
| Standard | Recombinant mouse Spp1 (Leu17–Asn294), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.8–7.5% (3 samples, n = 16 each) |
| Inter-assay CV | 4.9–9.0% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:50. In the supplier’s tests, serum and plasma samples measured 44–168 ng/mL.
Example standard curve (OD450, pg/mL): 0 = 0.025; 156 = 0.065; 312 = 0.103; 625 = 0.178; 1250 = 0.314; 2500 = 0.572; 5000 = 1.122; 10000 = 1.85. 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. | 0.025 | 0.065 | 0.103 | 0.178 | 0.314 | 0.572 | 1.122 | 1.85 |
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) | 306 | 1986 | 5401 | 308 | 2000 | 4959 |
| Standard deviation | 14.68 | 117.17 | 405.07 | 15.09 | 142 | 447.11 |
| CV (%) | 4.8% | 7.5% | 7.5% | 4.9% | 7.1% | 9% |
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.
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
Hua-Mo Yin et al. (2019) Surface Epitaxial Crystallization-Directed Nanotopography for Accelerating Preosteoblast Proliferation and Osteogenic Differentiation. ACS Applied Materials & Interfaces. 10.1021/acsami.9b14800
Min Ma et al. (2022) Baohuoside I Inhibits Osteoclastogenesis and Protects Against Ovariectomy-Induced Bone Loss. Frontiers in Pharmacology. 10.3389/fphar.2022.874952
Min-Xuan Xu et al. (2018) Activated iRhom2 drives prolonged PM2.5 exposure-triggered renal injury in Nrf2-defective mice. Nanotoxicology. 10.1080/17435390.2018.1513093
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
Hua-Mo Yin et al. (2019) Nanotopographical polymeric surface with mussel-inspired decoration to enhance osteoblast differentiation. Applied Surface Science. 10.1016/j.apsusc.2019.03.193
Baisong Zhao et al. (2018) Promoting osteoblast proliferation on polymer bone substitutes with bone-like structure by combining hydroxyapatite and bioactive glass. Materials Science & Engineering C-Materials for Biological Applications. 10.1016/j.msec.2018.11.006
Chen Liu et al. (2025) Osteopontin aggravates myocardial fibrosis by promoting phenotypic transition of cardiac fibroblasts via Hippo-YAP pathway. Biochimica et Biophysica ACTA-Molecular Basis of Disease. 10.1016/j.bbadis.2025.168150
Wentao Liu et al. (2025) Pro-Atherosclerotic Effects of Osteopontin Is Contributed to Promoting Foam Cell Formation Derived From VSMCs by Inhibiting Cholesterol Efflux. FASEB Journal. 10.1096/fj.202403104RR
Pan Liu et al. (2025) Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling. iScience. 10.1016/j.isci.2025.112107
