Human Osteopontin (OPN) ELISA

RayBio® Human Osteopontin (SPP1) ELISA Kit for cell culture supernatants, plasma, and serum samples.

Lead time: Typically ships within 1-2 business days. No Friday shipments.


Assay Format

Detection Range:
50 pg/ml - 18000 pg/ml
Recommended Dilution (serum & plasma):
3 - 100 fold
50 pg/ml

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This ELISA kit shows no cross-reactivity with any of the following cytokines tested (human Angiogenin, BDNF, BLC, ENA-78, FGF- 4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin, MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Number of Targets Detected:
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

1, 2, or 5 x 96-Well Strip Microplate Kit

Protein Information

Accession Number:
  • P10451
Gene ID:
  • 6696
Gene Symbols:
  • BNSP
  • OPN
  • PSEC0156
  • SPP1
Protein Name & Synonyms:
Osteopontin (Bone sialoprotein 1) (Nephropontin) (Secreted phosphoprotein 1) (SPP-1) (Urinary stone protein) (Uropontin)
Target Species:
  • Human

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

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The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Wu T., Ding H., Han J., et al. Antibody-Array-Based Proteomic Screening of Serum Markers in Systemic Lupus Erythematosus: A Discovery Study. J Proteome Res. 2016 Jul 1;15(7):2102-14. doi: 10.1021/acs.jproteome.5b00905.
    Species: Human
    Sample type: Serum (Lupus Nephritis patients)
  2. Nagy E, Eriksson P, Yousry M, et al. Valvular osteoclasts in calcification and aortic valve stenosis severity. Int J Cardiol. 2013 Feb 27; [Epub ahead of print] doi:10.1016/j.ijcard.2013.01.207.
    Species: Human
    Sample type: Plasma (intra-and interassay coefficients of variation were < 10% and < 12%)
  3. Bassyouni IH, El-Wakd MM, Bassyouni RH. Soluble levels of osteopontin in patients with Behcet’s disease: association with disease activity and vascular involvement. J Clin Immunol. 2013 Feb;33(2):361-367.
    Species: Human
    Sample type: Plasma
  4. Bassyouni I., Bassyouni R., Ibrahim N., Soliman A. Elevated serum osteopontin levels in chronic hepatitis C virus infection: association with autoimmune rheumatologic manifestations. J Clin Immunol. 2012 Dec;32(6):1262-9. doi: 10.1007/s10875-012-9727-7.
    Species: Human
    Sample type: Serum (Chronic Hep C patients in autoimmune rheumatologic manifestations)
  5. Von Neuhoff N, Oumeraci T, Wolf T, et al. Monitoring CSF Proteome Alterations in Amyotrophic Lateral Sclerosis: Obstacles and Perspectives in Translating a Novel Marker Panel to the Clinic. Borchelt DR, ed. PLoS ONE. 2012;7(9):e44401. doi:10.1371/journal.pone.0044401.
    Species: Human
    Sample type: CSF (ALS patients)
  6. Taranta-Janusz K., Wasilewska A., Stypulkowska J., Sutula M. Osteopontin and symmetric dimethylarginine plasma levels in solitary functioning kidney in children. Acta Paediatr. 2012 Aug;101(8):e369-72. doi: 10.1111/j.1651-2227.2012.02690.x
    Species: Human
    Sample type: Plasma (solitary functioning kidney patient samples)
  7. Gluba-Brzozka A., et al. Markers of increased cardiovascular risk in patients with chronic kidney disease. Lipids in Health and Disease 2014, 13:135 doi:10.1186/1476-511X-13-135
    Species: Human
    Sample type: Serum
  8. Erturkler, Enis, et al. "Plasma osteopontin levels in patients with Behcet's disease and psoriasis." European Journal of Dermatology 21.2 (2011): 203-208.
    Species: Human
    Sample type: Plasma
  9. Shaker, Olfat, et al. "Osteopontin gene polymorphisms as predictors for the efficacy of interferon therapy in chronic hepatitis C Egyptian patients with genotype 4." Cell biochemistry and function 31.7 (2013): 620-625.
    Species: Human
    Sample type: Serum (Chronic Hepatitis C patients)
  10. Acar A., Cevik M., Arikanoglu A., et al. Serum levels of calcification inhibitors in patients with intracerebral hemorrhage. Int J Neurosci. 2012 May;122(5):227-32. doi: 10.3109/00207454.2011.642039.
    Species: Human
    Sample type: Serum (Intracerebral Hemorrhage patients)

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This product is furnished for LABORATORY RESEARCH USE ONLY.
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