Human Osteopontin (OPN) Quantikine ELISA Kit

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Human Osteopontin/OPN Standard Curve
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Citations (38)
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Human Osteopontin (OPN) Quantikine ELISA Kit Summary

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.

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.

Precision

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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 2.3 4.9 9.3 2.36 4.81 9.17
Standard Deviation 0.092 0.126 0.267 0.155 0.274 0.499
CV% 4 2.6 2.9 6.6 5.7 5.4

Recovery

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

Linearity

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 ELISA Linearity

Data Examples

Human Osteopontin/OPN Standard Curve

Product Datasheets

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
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

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.

Citations for Human Osteopontin (OPN) Quantikine ELISA Kit

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.

38 Citations: Showing 1 - 10
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  1. Upregulation of osteopontin expression via the interaction of macrophages and fibroblasts under IL-1b stimulation
    Authors: T Shimodaira, K Matsuda, T Uchibori, M Sugano, T Uehara, T Honda
    Cytokine, 2018;110(0):63-69.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. Association of plasma osteopontin with diabetic retinopathy in Asians with type 2 diabetes
    Authors: X Zhang, WK Chee, S Liu, S Tavinthara, CF Sum, SC Lim, N Kumari
    Mol. Vis., 2018;24(0):165-173.
    Species: Human
    Sample Types: Plasma
  3. Methylglyoxal-bis-guanylhydrazone inhibits osteopontin expression and differentiation in cultured human monocytes
    Authors: X Jin, H Xu, MS McGrath
    PLoS ONE, 2018;13(3):e0192680.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Effects of oral paricalcitol therapy on arterial stiffness and osteopontin in hypertensive patients with chronic kidney disease and secondary hyperparathyroidism
    Authors: M Giakoumis, C Tsioufis, K Dimitriadi, M Sonikian, A Kasiakogia, E Andrikou, T Kalos, D Konstantin, K Filis, D Petras, D Tousoulis
    Hellenic J Cardiol, 2018;0(0):.
    Species: Human
    Sample Types: Blood
  5. 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 Types: Cell Culture Supernates
  6. Epigallocatechin-3-Gallate Upregulates miR-221 to Inhibit Osteopontin-Dependent Hepatic Fibrosis
    PLoS ONE, 2016;11(12):e0167435.
  7. Development of an in vitro human liver system for interrogating nonalcoholic steatohepatitis
    JCI Insight, 2016;1(20):e90954.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. IL-6 trans-signaling is another pathway to upregulate Osteopontin
    Cytokine, 2016;90(0):88-95.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Osteopontin is a multi-faceted pro-tumorigenic driver for central nervous system lymphoma
    Oncotarget, 2016;7(22):32156-71.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. 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 Types: Plasma
  11. 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 Types: Cell Culture Supernates
  12. 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 Types: Serum
  13. 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 Types: Pleural Fluid
  14. 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 Types: Plasma
  15. 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 Types: Serum
  16. 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 Types: Tissue Homogenates
  17. 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 Types: Serum
  18. 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 Types: Urine
  19. 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 Types: Plasma
  20. 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 Types: Plasma
  21. 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 Types: Plasma
  22. 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 Types: Serum
  23. An in vivo platform for tumor biomarker assessment.
    Authors: Servais EL, Suzuki K, Colovos C
    PLoS ONE, 2011;6(10):e26722.
    Species: Human
    Sample Types: 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 Types: Plasma
  25. Osteopontin is expressed and functional in human eosinophils.
    Authors: Puxeddu I, Berkman N, Ribatti D
    Allergy, 2010;65(2):168-74.
    Species: Human
    Sample Types: BALF
  26. 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 Types: Plasma
  27. 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 Types: Plasma
  28. 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 Types: Serum
  29. 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 Types: Cell Culture Supernates
  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 Types: Cell Culture Supernates
  31. 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 Types: Plasma
  32. 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 Types: Plasma
  33. 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 Types: Cell Culture Supernates
  34. 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 Types: Plasma
  35. 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 Types: Cell Culture Supernates
  36. 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 Types: Cell Culture Supernates
  37. 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 Types: Serum
  38. 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 Types: Serum

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Human Osteopontin (OPN) Quantikine ELISA Kit
By Anonymous on 12/01/2017
Sample Tested: Human cell conditioned medium