| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Alternative Names | SPP1; BNSP; BSPI; ETA-1; MGC110940; OPN; SPP1/CALPHA1 fusion; bone sialoprotein I; early T-lymphocyte activation 1; osteopontin; osteopontin/immunoglobulin alpha 1 heavy chain constant region fusion protein; s |
| Assay Time | |
| Assay Type | |
| Detection Method | |
| Gene ID | |
| Product Type | |
| Reactivity | |
| Sample Type(s) | Cell culture supernatants, Serum, Plasma, Other biological fluids |
| Shipping | |
| Storage |
Features & Benefits
Mouse Osteopontin (OPN) ELISA Kit has high sensitivity and excellent specificity for detection of Mouse SPP1. No significant cross-reactivity or interference between Mouse SPP1 and analogues was observed.
Background
Osteopontin (OPN), is a secreted multifunctional glyco-phosphoprotein with roles in bone metabolism, immune regulation, tissue remodeling , cell survival, and tumor progression . Gene structure and chromosomal location identify OPN as a member of the small integrin-binding ligand N-linked glycoprotein (SIBLING) family that also includes BSP,BMP1,DSPP, ENAM and MEPE. Murine OPN issynthesized as a 294 amino acid (aa) precursor protein with a predicted 16 aa signal peptide and a highly unusual mature protein sequence, containing 68 acidic aa and 23 potential Ser/Thr phosphorylation sites. Although the predicted molecular weight of OPN is 31 kDa,phosphorylation and N- and O-glycosylation may allow it to appear as large as 75 kDa. Variability in post-translational modifications can influence the activity of OPN.
This Osteopontin (OPN) ELISA kit is validated for use with Cell culture supernatants, Serum, Plasma, Other biological fluids. Samples should be collected, processed, and stored correctly to preserve analyte integrity — avoid repeated freeze-thaw cycles and centrifuge to remove particulates before use. Dilute samples exceeding the kit's detection range using the supplied assay diluent. Hemolytic, icteric, or lipemic samples may affect assay performance and should be tested with caution.
This is a sandwich ELISA kit employing an HRP (horseradish peroxidase)-conjugated secondary antibody paired with a TMB (3,3′,5,5′-tetramethylbenzidine) colorimetric substrate. In the sandwich format, the target analyte Osteopontin (OPN) captured on the microplate surface is detected by the conjugated antibody, generating a colorimetric signal proportional to analyte concentration. The reaction is stopped and absorbance measured at 450 nm on a standard microplate reader.
The complete protocol, from sample addition to final plate reading, requires approximately 3–5 hours. This includes two incubation periods (analyte binding and detection antibody steps), intermediate wash cycles to remove unbound material, 15–30 minutes of TMB substrate development, and final stop-solution addition before absorbance reading. Exact timing will vary with experience level and the number of samples processed in parallel.
Required equipment: (1) a microplate spectrophotometer capable of reading absorbance at 450 nm (reference wavelength 540–570 nm recommended for background correction); (2) precision single-channel or multichannel pipettes; (3) a plate washer or multichannel aspirator; (4) a microcentrifuge for sample clarification; and (5) a 37°C incubator or stable room-temperature environment. No fluorescence or luminescence reader is required — standard colorimetric plate readers are fully compatible with this kit.
This ELISA kit is formally validated for Mouse. Cross-reactivity with species not listed in the specification has not been independently characterized. Variability in protein sequence homology across species means that performance in unlisted species cannot be guaranteed without additional validation. For cross-species detection requirements or non-standard sample matrices, please contact BioHippo support or refer to the manufacturer's technical team for guidance.
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