Mouse & Rat Osteopontin (OPN) ELISA Kit - Quantikine

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Mouse/Rat Osteopontin/OPN ELISA Standard Curve
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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 (pg/mL) 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

Scientific Data

Mouse/Rat Osteopontin/OPN ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
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
&#9888; 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.

45 Citations: Showing 1 - 10
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  1. Use of Local Melatonin with Xenogeneic Bone Graft to Treat Critical-Size Bone Defects in Rats with Osteoporosis: A Randomized Study
    Authors: Costa, KLD;Abreu, LF;Tolomei, CB;Eleutério, RG;Basting, R;Balbinot, G;Collares, FM;Lopes, P;Veiga, N;Fernandes, GVO;Peruzzo, DC;
    Journal of functional biomaterials
    Species: Rat
    Sample Types: Cell Culture Supernates
  2. Tumor cell derived osteopontin and prostaglandin E2 synergistically promote the expansion of myeloid derived suppressor cells during the tumor immune escape phase
    Authors: Mamand, DR;Bazaz, S;Mohammad, DK;Saher, O;Wiklander, OPB;Sadeghi, B;Hassan, M;El-Andaloussi, S;Abedi-Valugerdi, M;
    International immunopharmacology
    Species: Mouse
    Sample Types: Serum, Cell Culture Supernates
  3. Next-Generation SINE Compound KPT-8602 Ameliorates Dystrophic Pathology in Zebrafish and Mouse Models of DMD
    Authors: KG English, AL Reid, A Samani, GJF Coulis, SA Villalta, CJ Walker, S Tamir, MS Alexander
    Biomedicines, 2022-09-26;10(10):.
    Species: Mouse
    Sample Types: Serum
  4. Evaluation of renal injury and function biomarkers, including symmetric dimethylarginine (SDMA), in the rat passive Heymann nephritis (PHN) model
    Authors: MJ Coyne, AE Schultze, DJ McCrann, RE Murphy, J Cross, M Strong-Tow, C Drake, R Mack
    PLoS ONE, 2022-05-27;17(5):e0269085.
    Species: Rat
    Sample Types: Urine
  5. Dysregulated Autophagy and Mitophagy in a Mouse Model of Duchenne Muscular Dystrophy Remain Unchanged Following Heme Oxygenase-1 Knockout
    Authors: O Mucha, K Kaziród, P Podkalicka, K Rusin, J Dulak, A ?oboda
    International Journal of Molecular Sciences, 2021-12-31;23(1):.
    Species: Mouse
    Sample Types: Serum
  6. Lipid-injured hepatocytes release sOPN to improve macrophage migration via CD44 engagement and pFak-NF&kappaB signaling
    Authors: X Jiang, F Zhang, X Ji, F Dong, H Yu, M Xue, Y Qiu, F Yang, X Hu, Z Bao
    Cytokine, 2021-02-27;142(0):155474.
    Species: Human, Mouse
    Sample Types: Cell Culture Supernates, Serum
  7. Praziquantel pharmacotherapy reduces systemic osteopontin levels and liver collagen content in murine schistosomiasis mansoni
    Authors: TA Pereira, G Vaz de Mel, E Trindade S, FE Pereira, M Maria de S
    International journal for parasitology, 2021-01-23;0(0):.
    Species: Mouse
    Sample Types: Serum
  8. Early Reciprocal Effects in a Murine Model of Traumatic Brain Injury and Femoral Fracture
    Authors: K Ritter, K Jung, C Dolderer, D Appel, CC Oswald, U Ritz, MKE Schäfer
    Mediators of Inflammation, 2021-01-18;2021(0):8835730.
    Species: Mouse
    Sample Types: Plasma
  9. Evaluation of urinary biomarkers for early detection of acute kidney injury in a rat nephropathy model
    Authors: K Kohl, E Herzog, G Dickneite, S Pestel
    J Pharmacol Toxicol Methods, 2020-08-07;0(0):106901.
    Species: Rat
    Sample Types: Urine
  10. Glycation and Serum Albumin Infiltration Contribute to the Structural Degeneration of Bioprosthetic Heart Valves
    Authors: A Frasca, Y Xue, AP Kossar, S Keeney, C Rock, A Zakharchen, M Streeter, RC Gorman, JB Grau, I George, JE Bavaria, A Krieger, DA Spiegel, RJ Levy, G Ferrari
    JACC Basic Transl Sci, 2020-08-05;5(8):755-766.
    Species: Rat
    Sample Types: Plasma
  11. Liver osteopontin is required to prevent the progression of age-related nonalcoholic fatty liver disease
    Authors: B Gómez-Sant, D Saenz de U, M Nuñez-Garc, F Gonzalez-R, X Buque, I Aurrekoetx, V Gutiérrez, MJ Gonzalez-R, C García-Mon, Á González-R, L Mosteiro, G Errazti, P Mifsut, S Gaztambide, L Castaño, C Martin, R Nogueiras, ML Martinez-C, WK Syn, P Aspichueta
    Aging Cell, 2020-07-07;0(0):e13183.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  12. Enhancer and super-enhancer dynamics in repair after ischemic acute kidney injury
    Authors: J Wilflingse, M Willi, HK Lee, H Olauson, J Jankowski, T Ichimura, R Erben, MT Valerius, L Hennighaus, JV Bonventre
    Nat Commun, 2020-07-07;11(1):3383.
    Species: Mouse
    Sample Types: Plasma
  13. The effects of macrophages on cardiomyocyte calcium-handling function using in vitro culture models
    Authors: PG Hitscheric, LH Xie, D Del Re, EJ Lee
    Physiol Rep, 2019-07-01;7(13):e14137.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  14. Pre-clinical evaluation of Minnelide as a therapy for acute myeloid leukemia
    Authors: B Giri, VK Gupta, B Yaffe, S Modi, P Roy, V Sethi, SP Lavania, SM Vickers, V Dudeja, S Banerjee, J Watts, A Saluja
    J Transl Med, 2019-05-20;17(1):163.
    Species: Human
    Sample Types: Whole Cells
  15. Osteopontin in the host response to Leishmania amazonensis
    Authors: E Giraud, E Rouault, L Fiette, JH Colle, D Smirlis, E Melanitou
    BMC Microbiol., 2019-02-08;19(1):32.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  16. Osteopontin potentiates PM-induced IL-1? and IL-1? production via the ERK/JNK signaling pathway
    Authors: J Wang, J Huang, M Zhu, S Chen, C Chen, C Miao, H He, Y Song
    Ecotoxicol. Environ. Saf., 2019-01-09;171(0):467-474.
    Species: Mouse
    Sample Types: Serum
  17. 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., 2018-11-05;50(11):142.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  18. 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., 2018-11-05;0(0):.
    Species: Mouse
    Sample Types: Serum
  19. 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, 2018-10-11;8(1):15130.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  20. 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, 2018-10-02;8(1):14626.
    Species: Mouse
    Sample Types: Serum
  21. Disruption of renal arginine metabolism promotes kidney injury in hepatorenal syndrome
    Authors: ZV Varga, K Erdelyi, J Paloczi, R Cinar, ZK Zsengeller, T Jourdan, C Matyas, NT Balazs, A Guillot, X Xiang, A Mehal, G Hasko, IE Stillman, S Rosen, B Gao, G Kunos, P Pacher
    Hepatology, 2018-10-01;0(0):.
    Species: Mouse
    Sample Types: Serum
  22. 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., 2018-04-25;0(0):.
    Species: Mouse
    Sample Types: Plasma
  23. Loss of NF-?B1 Causes Gastric Cancer with Aberrant Inflammation and Expression of Immune Checkpoint Regulators in a STAT-1-Dependent Manner
    Authors: LA O'Reilly, TL Putoczki, LA Mielke, JT Low, A Lin, A Preaudet, MJ Herold, K Yaprianto, L Tai, A Kueh, G Pacini, RL Ferrero, R Gugasyan, Y Hu, M Christie, S Wilcox, R Grumont, MDW Griffin, L O'Connor, GK Smyth, M Ernst, P Waring, S Gerondakis, A Strasser
    Immunity, 2018-03-20;48(3):570-583.e8.
    Species: Mouse
    Sample Types: Serum
  24. In vivo inhibition of nuclear factor of activated T-cells leads to atherosclerotic plaque regression in IGF-II/LDLR-/-ApoB100/100mice
    Authors: F Blanco, SE Heinonen, E Gurzeler, LM Berglund, AM Dutius And, O Kotova, AC Jönsson-Ry, S Ylä-Herttu, MF Gomez
    Diab Vasc Dis Res, 2018-03-02;0(0):1479164118759.
    Species: Mouse
    Sample Types: Plasma
  25. 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, 2018-02-22;3(4):.
    Species: Rat
    Sample Types: Serum
  26. 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, 2017-10-12;7(1):13084.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  27. 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, 2017-07-26;7(1):6496.
    Species: Mouse
    Sample Types: Plasma
  28. 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, 2017-05-12;12(5):e0177640.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  29. Deficient TSC1/TSC2-complex suppression of SOX9-osteopontin-AKT signalling cascade constrains tumour growth in tuberous sclerosis complex
    Authors: Fuquan Jin
    Hum. Mol. Genet, 2017-01-15;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  30. Ultrastructural changes during osteogenic differentiation in mesenchymal stromal cells cultured in alginate hydrogel
    Authors: J Grzesiak, A ?mieszek, K Marycz
    Cell Biosci, 2017-01-03;7(0):2.
    Species: Rat
    Sample Types: Cell Culture Supernates
  31. The P2X4 receptor is required for neuroprotection via ischemic preconditioning
    Sci Rep, 2016-05-13;6(0):25893.
    Species: Mouse
    Sample Types: Tissue Homogenates
  32. Age-associated vascular inflammation promotes monocytosis during atherogenesis
    Aging Cell, 2016-05-02;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  33. 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, 2015-08-04;10(8):e0134764.
    Species: Rat
    Sample Types: Urine
  34. 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, 2015-03-31;2015(0):428473.
    Species: Mouse
    Sample Types: Plasma
  35. PHEX mimetic (SPR4-peptide) corrects and improves HYP and wild type mice energy-metabolism.
    Authors: Zelenchuk L, Hedge A, Rowe P
    PLoS ONE, 2014-05-19;9(5):e97326.
    Species: Mouse
    Sample Types: Serum
  36. 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, 2014-02-04;9(2):e88089.
    Species: Rat
    Sample Types: Cell Culture Supernates
  37. 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, 2014-01-01;184(1):159-70.
    Species: Mouse
    Sample Types: Tissue Homogenates
  38. 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, 2013-07-27;25(12):2362-73.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  39. 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., 2012-03-23;31(0):26.
    Species: Mouse
    Sample Types: Serum
  40. 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., 2010-12-06;25(3):968-78.
    Species: Rat
    Sample Types: Tissue Homogenates
  41. 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., 2010-09-15;285(47):37041-50.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  42. 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, 2010-01-27;59(4):935-46.
    Species: Mouse
    Sample Types: Plasma
  43. 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., 2009-05-28;16(8):982-9.
    Species: Mouse
    Sample Types: Serum
  44. Heterozygous disruption of the PTEN promotes intestinal neoplasia in APCmin/+ mouse: roles of osteopontin.
    Authors: Shao J, Washington MK, Saxena R, Sheng H
    Carcinogenesis, 2007-08-11;28(12):2476-83.
    Species: Rat
    Sample Types: Cell Culture Supernates
  45. 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., 2007-07-10;361(1):140-5.
    Species: Mouse
    Sample Types: Plasma

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Mouse/Rat Osteopontin (OPN) Quantikine ELISA Kit
By Anonymous on 01/16/2022
Sample Tested: Serum

Easy to perform.