| 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 EZ-Set™ kit supplies a matched capture and detection antibody pair and a standard for building your own sandwich ELISA for human osteopontin (gene SPP1) in cell culture supernatants, serum, plasma (heparin, EDTA), human milk and urine. You coat the plate with the capture antibody, add samples and standards, detect with the biotinylated antibody and an avidin–biotin–peroxidase complex, develop with TMB and read at 450 nm. Plates, diluents, wash buffer, TMB and stop solution are not included.
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
| Mouse anti-human OPN/Osteopontin monoclonal antibody (Capture Antibody) | 500 µL, 4 µg/mL (recommended dilution 1:100) |
|---|---|
| Biotinylated goat anti-human OPN/Osteopontin polyclonal antibody (Detection Antibody) | 500 µL (recommended dilution 1:100) |
| Avidin-Biotin-Peroxidase Complex (ABC) | 500 µL (recommended dilution 1:100) |
| Lyophilized recombinant human OPN/Osteopontin standard | 3 × 10 ng/tube |
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 |
| Standard | Recombinant human SPP1 (Ile17–Asn300), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
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. | 0.056 | 0.110 | 0.156 | 0.244 | 0.457 | 0.946 | 1.677 | 2.085 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0482-CA | Mouse anti-human OPN/Osteopontin monoclonal antibody (Capture Antibody) | 500 µl, 4 µg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ0482-DA | Biotinylated goat anti-human OPN/Osteopontin polyclonal antibody (Detection Antibody) | 500 µl (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EZ0482-ST | Lyophilized recombinant human OPN/Osteopontin standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
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.
