Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit

  (21 citations)     
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Product Details
Assay Procedure
Citations (21)
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  • 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.
 Osteopontin/OPN [HRP]
 Osteopontin/OPN [HRP]
Product Datasheets

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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. 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:

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.

21 Citations: Showing 1 - 10
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Species
Sample Type
  1. 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 Type: Plasma
  2. 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 Type: Serum
  3. 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 Type: Cell Culture Supernates
  4. 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 Type: Plasma
  5. 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 Type: Cell Culture Supernates
  6. Age-associated vascular inflammation promotes monocytosis during atherogenesis
    Aging Cell, 2016;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  7. 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 Type: Cell Culture Supernates
  8. The P2X4 receptor is required for neuroprotection via ischemic preconditioning
    Sci Rep, 2016;6(0):25893.
    Species: Mouse
    Sample Type: Tissue Homogenates
  9. 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 Type: Urine
  10. 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 Type: Plasma
  11. 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 Type: Tissue Homogenates
  12. 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 Type: Serum
  13. 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 Type: Cell Culture Supernates
  14. 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 Type: Cell Culture Supernates
  15. 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 Type: Serum
  16. 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 Type: Tissue Homogenates
  17. 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 Type: Cell Culture Supernates
  18. 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 Type: Plasma
  19. 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 Type: Serum
  20. 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 Type: Cell Culture Supernates
  21. 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 Type: Plasma
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