Human Osteopontin (OPN) Quantikine ELISA Kit

(34 citations)
(1 Review)
  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (50 uL), Human Milk (50 uL)
  • Sensitivity
    0.024 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Urine, Human Milk)
  • Specificity
    Natural and recombinant human Osteopontin
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC136, Quantikine Immunoassay Control Set 960 for Human Osteopontin - Please Inquire
Product Summary
The Quantikine Human Osteopontin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Osteopontin in cell culture supernates, plasma, urine, and human milk. It contains NS0-expressed recombinant human Osteopontin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Osteopontin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Osteopontin.

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Urine, Human Milk
Intra-Assay Precision Inter-Assay Precision
Standard Deviation0.0920.1260.2670.1550.2740.499


The recovery of Osteopontin spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 104 86-113
EDTA Plasma (n=4) 102 92-114
Heparin Plasma (n=4) 101 95-112
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Osteopontin were serially diluted with the Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Osteopontin (OPN) Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Osteopontin/OPN
Osteopontin (OPN), also known as bone sialoprotein (BSP), is a secreted SIBLING family protein that can be variably modified by O- and N-glycosylation, sulfation, phosphorylation, and transglutamination. OPN is widely expressed and is prominent in mineralized tissues. It inhibits bone mineralization and kidney stone formation and promotes inflammation, cell ad¬hesion, and migration. Its expression is upregulat¬ed during inflammation, obesity, atherosclerosis, cancer, and tissue damage and contributes to the pathophysiology of these conditions. The central region of OPN contains RGD and non-RGD binding sites for multiple integrins. Adjacent to the RGD motif is the sequence SLAYGLR (SVVYGLR in human) which serves as a cryptic binding site for additional integrins: it is masked in full length OPN but is exposed following OPN cleavage by multiple proteases in tumors and sites of tissue injury.
    • Long Name
      Secreted Phosphoprotein 1 [BNSP]
    • Entrez Gene IDs
      6696 (Human); 20750 (Mouse); 25353 (Rat); 281499 (Bovine);
    • Alternate Names
      BNSP; Bone sialoprotein 1; Eta-1; MGC110940; Nephropontin; OPN; Osteopontin; secreted phosphoprotein 1bone sialoprotein I, early T-lymphocyteactivation 1); secreted phosphoprotein-1 (osteopontin, bone sialoprotein); Spp1; SPP-1; SPP1/CALPHA1 fusion; Urinary stone protein; uropontin;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
    1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
    2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

    3. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    8. 200 µL Conjugate
    9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 4 times.

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    34 Citations: Showing 1 - 10
    Filter your results:

    Sample Type
    1. Osteopontin is Critical for Hyperactive mTOR-Induced Tumorigenesis in Oral Squamous Cell Carcinoma
      Authors: N Gan, S Zou, W Hang, D Yang, X Zhang, Y Yin
      J Cancer, 2017;8(8):1362-1370.
      Species: Human
      Sample Type: Cell Culture Supernates
    2. Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis
      JCI Insight, 2016;1(20):e90954.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. Epigallocatechin-3-Gallate Upregulates miR-221 to Inhibit Osteopontin-Dependent Hepatic Fibrosis
      PLoS ONE, 2016;11(12):e0167435.
    4. Targeting of the leukemia microenvironment by c(RGDfV) overcomes the resistance to chemotherapy in acute myeloid leukemia in biomimetic polystyrene scaffolds
      Oncol Lett, 2016;12(5):3278-3284.
      Species: Human
      Sample Type: Cell Culture Supernates
    5. IL-6 trans-signaling is another pathway to upregulate Osteopontin
      Cytokine, 2016;90(0):88-95.
      Species: Human
      Sample Type: Cell Culture Supernates
    6. Osteopontin is a multi-faceted pro-tumorigenic driver for central nervous system lymphoma
      Oncotarget, 2016;7(22):32156-71.
      Species: Human
      Sample Type: Cell Culture Supernates
    7. Immunomodulation Induced by Stem Cell Mobilization and Harvesting in Healthy Donors: Increased Systemic Osteopontin Levels after Treatment with Granulocyte Colony-Stimulating Factor
      Int J Mol Sci, 2016;17(7):.
      Species: Human
      Sample Type: Plasma
    8. Biomarkers of inflammation and axonal degeneration/damage in patients with newly diagnosed multiple sclerosis: contributions of the soluble CD163 CSF/serum ratio to a biomarker panel.
      Authors: Stilund M, Gjelstrup M, Petersen T, Moller H, Rasmussen P, Christensen T
      PLoS ONE, 2015;10(4):e0119681.
      Species: Human
      Sample Type: Serum
    9. Interleukin 4, interleukin 6 and osteopontin-serological markers of head and neck malignancy in primary diagnostics: A pilot study.
      Authors: Aderhold C, Grobschmidt G, Sauter A, Faber A, Hormann K, Schultz J
      Oncol Lett, 2014;8(3):1112-1118.
      Species: Human
      Sample Type: Serum
    10. Prognostic value of secreted phosphoprotein-1 in pleural effusion associated with non-small cell lung cancer.
      Authors: Zhang, He, Liu, Hong-bin, Yuan, Dong-mei, Wang, Zhao-fen, Wang, Yun-fen, Song, Yong
      BMC Cancer, 2014;14(0):280.
      Species: Human
      Sample Type: Pleural Fluid
    11. Prediction of cardiovascular events in statin-treated stable coronary patients of the treating to new targets randomized controlled trial by lipid and non-lipid biomarkers.
      Authors: Arsenault B, Barter P, DeMicco D, Bao W, Preston G, LaRosa J, Grundy S, Deedwania P, Greten H, Wenger N, Shepherd J, Waters D, Kastelein J
      PLoS ONE, 2014;9(12):e114519.
      Species: Human
      Sample Type: Plasma
    12. Thrombin-cleaved fragments of osteopontin are overexpressed in malignant glial tumors and provide a molecular niche with survival advantage.
      Authors: Yamaguchi Y, Shao Z, Sharif S, Du X, Myles T, Merchant M, Harsh G, Glantz M, Recht L, Morser J, Leung L
      J Biol Chem, 2013;288(5):3097-111.
      Species: Human
      Sample Type: Tissue Homogenates
    13. Osteopontin is a prognostic biomarker in non-small cell lung cancer.
      Authors: Rud A, Boye K, Oijordsbakken M, Lund-Iversen M, Halvorsen A, Solberg S, Berge G, Helland A, Brustugun O, Maelandsmo G
      BMC Cancer, 2013;13(0):540.
      Species: Human
      Sample Type: Serum
    14. Prognostic utility of pre-operative circulating osteopontin, carbonic anhydrase IX and CRP in renal cell carcinoma.
      Br. J. Cancer, 2012;107(7):1131-7.
      Species: Human
      Sample Type: Plasma
    15. Osteopontin is a prognostic factor for survival of acute myeloid leukemia patients.
      Authors: Liersch R, Gerss J, Schliemann C, Bayer M, Schwoppe C, Biermann C, Appelmann I, Kessler T, Lowenberg B, Buchner T, Hiddemann W, Muller-Tidow C, Berdel WE, Mesters R
      Blood, 2012;119(22):5215-20.
      Species: Human
      Sample Type: Plasma
    16. A multi-analyte assay for the non-invasive detection of bladder cancer.
      Authors: Goodison, Steve, Chang, Myron, Dai, Yunfeng, Urquidi, Virginia, Rosser, Charles
      PLoS ONE, 2012;7(10):e47469.
      Species: Human
      Sample Type: Urine
    17. Serum levels of beta-catenin as a potential marker for genotype 4/hepatitis C-associated hepatocellular carcinoma.
      Authors: Zekri A, Bahnassy A, Alam El-Din H, Morsy H, Shaarawy S, Moharram N, Daoud S
      Oncol Rep, 2011;26(4):825-31.
      Species: Human
      Sample Type: Serum
    18. An in vivo platform for tumor biomarker assessment.
      Authors: Servais EL, Suzuki K, Colovos C
      PLoS ONE, 2011;6(10):e26722.
      Species: Human
      Sample Type: Plasma
    19. Plasma markers for identifying patients with metastatic melanoma.
      Authors: Kluger HM, Hoyt K, Bacchiocchi A, Mayer T, Kirsch J, Kluger Y, Sznol M, Ariyan S, Molinaro A, Halaban R
      Clin. Cancer Res., 2011;17(8):2417-25.
      Species: Human
      Sample Type: Plasma
    20. Changes in osteopontin and in biomarkers of bone turnover during human endotoxemia.
      Authors: Grimm G, Vila G, Bieglmayer C
      Bone, 2010;47(2):388-91.
      Species: Human
      Sample Type: Plasma
    21. A multi-analyte serum test for the detection of non-small cell lung cancer.
      Authors: Farlow EC, Vercillo MS, Coon JS, Basu S, Kim AW, Faber LP, Warren WH, Bonomi P, Liptay MJ, Borgia JA
      Br. J. Cancer, 2010;103(8):1221-8.
      Species: Human
      Sample Type: Serum
    22. Osteopontin is expressed and functional in human eosinophils.
      Authors: Puxeddu I, Berkman N, Ribatti D
      Allergy, 2010;65(2):168-74.
      Species: Human
      Sample Type: BALF
    23. Circulating levels of osteopontin are closely related to glomerular filtration rate and cardiovascular risk markers in patients with chronic kidney disease.
      Authors: Lorenzen J, Kramer R, Kliem V, Bode-Boeger SM, Veldink H, Haller H, Fliser D, Kielstein JT
      Eur. J. Clin. Invest., 2010;40(4):294-300.
      Species: Human
      Sample Type: Plasma
    24. Osteopontin as a novel prognostic marker in stable ischaemic heart disease: a 3-year follow-up study.
      Authors: Georgiadou P, Iliodromitis EK, Kolokathis F, Varounis C, Gizas V, Mavroidis M, Capetanaki Y, Boudoulas H, Kremastinos DT
      Eur. J. Clin. Invest., 2010;0(0):.
      Species: Human
      Sample Type: Plasma
    25. Characterization of Gaucher disease bone marrow mesenchymal stromal cells reveals an altered inflammatory secretome.
      Authors: Campeau PM, Rafei M, Boivin MN, Sun Y, Grabowski GA, Galipeau J
      Blood, 2009;114(15):3181-90.
      Species: Human
      Sample Type: Cell Culture Supernates
    26. Abnormal cytokine production by bone marrow stromal cells of multiple myeloma patients in response to RPMI8226 myeloma cells.
      Authors: Zdzisinska B, Bojarska-Junak A, Dmoszynska A, Kandefer-Szerszen M
      Arch. Immunol. Ther. Exp. (Warsz.), 2008;56(3):207-21.
      Species: Human
      Sample Type: Cell Culture Supernates
    27. Activation of the osteopontin/matrix metalloproteinase-9 pathway correlates with prostate cancer progression.
      Authors: Castellano G, Malaponte G, Mazzarino MC, Figini M, Marchese F, Gangemi P, Travali S, Stivala F, Canevari S, Libra M
      Clin. Cancer Res., 2008;14(22):7470-80.
      Species: Human
      Sample Type: Plasma
    28. Abrogation of T cell quiescence characterizes patients at high risk for multiple sclerosis after the initial neurological event.
      Authors: Corvol JC, Pelletier D, Henry RG, Caillier SJ, Wang J, Pappas D, Casazza S, Okuda DT, Hauser SL, Oksenberg JR, Baranzini SE
      Proc. Natl. Acad. Sci. U.S.A., 2008;105(33):11839-44.
      Species: Human
      Sample Type: Plasma
    29. Intensive lipid-lowering therapy ameliorates novel calcification markers and GSM score in patients with carotid stenosis.
      Authors: Kadoglou NP, Gerasimidis T, Moumtzouoglou A, Kapelouzou A, Sailer N, Fotiadis G, Vitta I, Katinios A, Kougias P, Bandios S, Voliotis K, Karayannacos PE, Liapis CD
      Eur J Vasc Endovasc Surg, 2008;35(6):661-8.
      Species: Human
      Sample Type: Plasma
    30. Innate immune responses to TREM-1 activation: overlap, divergence, and positive and negative cross-talk with bacterial lipopolysaccharide.
      Authors: Dower K, Ellis DK, Saraf K, Jelinsky SA, Lin LL
      J. Immunol., 2008;180(5):3520-34.
      Species: Human
      Sample Type: Cell Culture Supernates
    31. Heterozygous disruption of the PTEN promotes intestinal neoplasia in APCmin/+ mouse: roles of osteopontin.
      Authors: Shao J, Washington MK, Saxena R, Sheng H
      Carcinogenesis, 2007;28(12):2476-83.
      Species: Human
      Sample Type: Cell Culture Supernates
    32. The new tumor-suppressor gene inhibitor of growth family member 4 (ING4) regulates the production of proangiogenic molecules by myeloma cells and suppresses hypoxia-inducible factor-1 alpha (HIF-1alpha) activity: involvement in myeloma-induced angiogenesis.
      Authors: Colla S, Tagliaferri S, Morandi F, Lunghi P, Donofrio G, Martorana D, Mancini C, Lazzaretti M, Mazzera L, Ravanetti L, Bonomini S, Ferrari L, Miranda C, Ladetto M, Neri TM, Neri A, Greco A, Mangoni M, Bonati A, Rizzoli V, Giuliani N
      Blood, 2007;110(13):4464-75.
      Species: Human
      Sample Type: Cell Culture Supernates
    33. An integrated clinical-genomics approach identifies a candidate multi-analyte blood test for serous ovarian carcinoma.
      Authors: Meinhold-Heerlein I, Bauerschlag D, Zhou Y, Sapinoso LM, Ching K, Frierson H, Brautigam K, Sehouli J, Stickeler E, Konsgen D, Hilpert F, von Kaisenberg CS, Pfisterer J, Bauknecht T, Jonat W, Arnold N, Hampton GM
      Clin. Cancer Res., 2007;13(2):458-66.
      Species: Human
      Sample Type: Serum
    34. Osteopontin expression and serum levels in metastatic uveal melanoma: a pilot study.
      Authors: Pe&amp;amp;apos;er J
      Invest. Ophthalmol. Vis. Sci., 2006;47(3):802-6.
      Species: Human
      Sample Type: Serum
    Expand to show all 34 Citations
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    Quantikine Immunoassay Control Set 960 for Human Osteopontin

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    Sample TestedHuman cell conditioned medium

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