Human Osteopontin (OPN) ELISA

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


Antigen 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

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

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
  2. Filis K., Martinakis V., Galyfos G., Sigala F., Theodorou D., Andreadou I., Zografos G. OSTEOPONTIN AND OSTEOPROTEGERIN AS POTENTIAL BIOMARKERS IN ABDOMINAL AORTIC ANEURYSM BEFORE AND AFTER TREATMENT. International Scholarly Research Notices, 2014 May, Epub Ahead of Print
    Species: Human
    Sample type: Serum
  3. 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
  4. Hussein Y., Alhazmi A., Alzahrani S., et al. Osteopontin as a Marker for Response to Pegylated Interferon Alpha-2b Treatment in Chronic HCV Saudi Patients. Vol 21, No. 12;Dec 2014. 445 - 454
    Species: Human
    Sample type: Serum
  5. 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
  6. Hafez SS, El Sebai AA, Al Aziz MM, et al. Alfa-Fetoprotein L3 Subfraction and Osteopontin: Novel Markers for the Diagnosis of Hepatocellular Carcinoma. J Am Sci. 2013;9(3).
    Species: Human
    Sample type: Serum
  7. 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
  8. 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
  9. Hydzik, P., D. Szpak, and E. Gomó?ka. "Osteopontin level in the serum of patients with alcoholic liver disease, treated for alcohol withdrawal syndrome--a preliminary report."?Przeglad lekarski?70.8 (2013): 542-545.
    Species: Human
    Sample type: Serum
  10. Sakr, Hala Al Sayed Ahmed, Laila Ateif Ahmed, and Wafaa Mohie?Eldeen Abdel Fattah. "Molecular Markers Predicting the Efficacy of Interferon Based Therapies in Patients with Chronic Hepatitis C."?Journal of American Science?7 (2013): 9.
    Species: Human
    Sample type: Serum

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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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