Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit

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Osteopontin/OPN ELISA Kit ELISA
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Mouse/Rat 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), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (50 uL)
Sensitivity
8.5 pg/mL
Assay Range
39.0 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant mouse and rat OPN. This assay cross-reacts 8.2% with recombinant Human Osteopontin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Mouse OPN Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse OPN in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse OPN and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse OPN 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 mouse OPN.

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, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 62 67 59
Mean 96 236 1085 116 279 1076
Standard Deviation 7.3 13.8 43.4 9.8 22.5 90
CV% 7.6 5.8 4 8.4 8.1 8.4

Recovery

The recovery of mouse OPN spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=7) 98 87-106
Rat Cell Culture Supernates (n=7) 93 82-107

Linearity

To assess the linearity of the assay, samples containing mouse or rat OPN were diluted with Calibrator Diluent and then assayed.
Mouse/Rat Osteopontin/OPN ELISA Linearity
Mouse/Rat Osteopontin/OPN ELISA Linearity

Data Examples

Mouse/Rat Osteopontin/OPN ELISA 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. 50 µL Assay Diluent
  4.   Add 50 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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 Mouse/Rat 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.

26 Citations: Showing 1 - 10
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  1. An osteopontin/CD44 immune checkpoint controls CD8+ T cell activation and tumor immune evasion
    Authors: JD Klement, AV Paschall, PS Redd, ML Ibrahim, C Lu, D Yang, E Celis, SI Abrams, K Ozato, K Liu
    J. Clin. Invest., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  2. Tumor-independent host secretomes induced by angiogenesis and immune-checkpoint inhibitors
    Authors: M Mastri, CR Lee, A Tracz, RS Kerbel, M Dolan, Y Shi, JML Ebos
    Mol. Cancer Ther., 2018;0(0):.
    Species: Mouse
    Sample Types: Plasma
  3. Nitric oxide-sensitive guanylyl cyclase stimulation improves experimental heart failure with preserved ejection fraction
    Authors: N Wilck, L Markó, A Balogh, K Kräker, F Herse, H Bartolomae, IA Szijártó, M Gollasch, N Reichhart, O Strauss, A Heuser, D Brockschni, A Kretschmer, R Lesche, F Sohler, JP Stasch, P Sandner, FC Luft, DN Müller, R Dechend, N Haase
    JCI Insight, 2018;3(4):.
    Species: Rat
    Sample Types: Serum
  4. Elevated extracellular calcium ions promote proliferation and migration of mesenchymal stem cells via increasing osteopontin expression
    Authors: MN Lee, HS Hwang, SH Oh, A Roshanzade, JW Kim, JH Song, ES Kim, JT Koh
    Exp. Mol. Med., 2018;50(11):142.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Atorvastatin provides a new lipidome improving early regeneration after partial hepatectomy in osteopontin deficient mice
    Authors: M Nuñez-Garc, B Gomez-Sant, D Saenz de U, D Mestre, F Gonzalez-R, X Buque, V Gutiérrez-, ML Martinez-C, WK Syn, O Fresnedo, P Aspichueta
    Sci Rep, 2018;8(1):14626.
    Species: Mouse
    Sample Types: Serum
  6. Multi-compartment scaffold fabricated via 3D-printing as in vitro co-culture osteogenic model
    Authors: E De Giglio, MA Bonifacio, AM Ferreira, S Cometa, ZY Ti, A Stanzione, K Dalgarno, P Gentile
    Sci Rep, 2018;8(1):15130.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  7. Uremia does not affect neointima formation in mice
    Authors: A Aarup, CH Nielsen, LS Bisgaard, I Bot, HH El-Ali, A Kjaer, LB Nielsen, TX Pedersen
    Sci Rep, 2017;7(1):6496.
    Species: Mouse
    Sample Types: Plasma
  8. Ultrastructural changes during osteogenic differentiation in mesenchymal stromal cells cultured in alginate hydrogel
    Authors: J Grzesiak, A ?mieszek, K Marycz
    Cell Biosci, 2017;7(0):2.
    Species: Rat
    Sample Types: Cell Culture Supernates
  9. Involvement of the accumbal osteopontin-interacting transmembrane protein 168 in methamphetamine-induced place preference and hyperlocomotion in mice
    Authors: K Fu, Y Miyamoto, K Otake, K Sumi, E Saika, S Matsumura, N Sato, Y Ueno, S Seo, K Uno, SI Muramatsu, A Nitta
    Sci Rep, 2017;7(1):13084.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Soluble bone-derived osteopontin promotes migration and stem-like behavior of breast cancer cells
    Authors: GM Pio, Y Xia, MM Piaseczny, JE Chu, AL Allan
    PLoS ONE, 2017;12(5):e0177640.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  11. Deficient TSC1/TSC2-complex suppression of SOX9-osteopontin-AKT signalling cascade constrains tumour growth in tuberous sclerosis complex
    Authors: Fuquan Jin
    Hum. Mol. Genet, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Age-associated vascular inflammation promotes monocytosis during atherogenesis
    Aging Cell, 2016;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  13. The P2X4 receptor is required for neuroprotection via ischemic preconditioning
    Sci Rep, 2016;6(0):25893.
    Species: Mouse
    Sample Types: Tissue Homogenates
  14. High Sodium-Induced Oxidative Stress and Poor Anticrystallization Defense Aggravate Calcium Oxalate Crystal Formation in Rat Hyperoxaluric Kidneys.
    Authors: Huang H, Ma M
    PLoS ONE, 2015;10(8):e0134764.
    Species: Rat
    Sample Types: Urine
  15. Nuclear factor of activated T cells is activated in the endothelium of retinal microvessels in diabetic mice.
    Authors: Zetterqvist A, Blanco F, Ohman J, Kotova O, Berglund L, de Frutos Garcia S, Al-Naemi R, Wigren M, McGuire P, Gonzalez Bosc L, Gomez M
    J Diabetes Res, 2015;2015(0):428473.
    Species: Mouse
    Sample Types: Plasma
  16. PHEX mimetic (SPR4-peptide) corrects and improves HYP and wild type mice energy-metabolism.
    Authors: Zelenchuk L, Hedge A, Rowe P
    PLoS ONE, 2014;9(5):e97326.
    Species: Mouse
    Sample Types: Serum
  17. The level of hepatic ABCC6 expression determines the severity of calcification after cardiac injury.
    Authors: Brampton C, Aherrahrou Z, Chen L, Martin L, Bergen A, Gorgels T, Erdmann J, Schunkert H, Szabo Z, Varadi A, Le Saux O
    Am J Pathol, 2014;184(1):159-70.
    Species: Mouse
    Sample Types: Tissue Homogenates
  18. Phenotypic modulation of primary vascular smooth muscle cells by short-term culture on micropatterned substrate.
    Authors: Chang S, Song S, Lee J, Yoon J, Park J, Choi S, Park J, Choi K, Choi C
    PLoS ONE, 2014;9(2):e88089.
    Species: Rat
    Sample Types: Cell Culture Supernates
  19. Plasma membrane Pdia3 and VDR interact to elicit rapid responses to 1alpha,25(OH)(2)D(3).
    Authors: Chen J, Doroudi M, Cheung J, Grozier A, Schwartz Z, Boyan B
    Cell Signal, 2013;25(12):2362-73.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  20. Osteopontin induces growth of metastatic tumors in a preclinical model of non-small lung cancer.
    Authors: Shojaei F, Scott N, Kang X, Lappin PB, Fitzgerald AA, Karlicek S, Simmons BH, Wu A, Lee JH, Bergqvist S, Kraynov E
    J. Exp. Clin. Cancer Res., 2012;31(0):26.
    Species: Mouse
    Sample Types: Serum
  21. Mineralocorticoid receptor-mediated DNA damage in kidneys of DOCA-salt hypertensive rats.
    Authors: Schupp N, Kolkhof P, Queisser N, Gartner S, Schmid U, Kretschmer A, Hartmann E, Oli RG, Schafer S, Stopper H
    FASEB J., 2011;25(3):968-78.
    Species: Rat
    Sample Types: Tissue Homogenates
  22. Protein-disulfide isomerase-associated 3 (Pdia3) mediates the membrane response to 1,25-dihydroxyvitamin D3 in osteoblasts.
    Authors: Chen J, Olivares-Navarrete R, Wang Y
    J. Biol. Chem., 2010;285(47):37041-50.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  23. Neutralization of osteopontin inhibits obesity-induced inflammation and insulin resistance.
    Authors: Kiefer FW, Zeyda M, Gollinger K, Pfau B, Neuhofer A, Weichhart T, Saemann MD, Geyeregger R, Schlederer M, Kenner L, Stulnig TM
    Diabetes, 2010;59(4):935-46.
    Species: Mouse
    Sample Types: Plasma
  24. Therapeutic silencing of an endogenous gene by siRNA cream in an arthritis model mouse.
    Authors: Takanashi M, Oikawa K, Sudo K, Tanaka M, Fujita K, Ishikawa A, Nakae S, Kaspar RL, Matsuzaki M, Kudo M, Kuroda M
    Gene Ther., 2009;16(8):982-9.
    Species: Mouse
    Sample Types: Serum
  25. LPS-induced biomarkers in mice: a potential model for identifying insulin sensitizers.
    Authors: Briscoe CP, Looper D, Tran P, Herrera J, McDonnell SR, Bhat BG
    Biochem. Biophys. Res. Commun., 2007;361(1):140-5.
    Species: Mouse
    Sample Types: Plasma
  26. 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: Rat
    Sample Types: Cell Culture Supernates

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