Mouse Osteoprotegerin/TNFRSF11B Quantikine ELISA Kit

(32 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), Serum (10 uL), EDTA Plasma (10 uL)
  • Sensitivity
    6.9 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse OPG
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Product Summary
The Quantikine Mouse OPG Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse OPG in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse OPG and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate recombinant mouse OPG accurately. Results obtained using natural mouse OPG showed dose response 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 natural mouse OPG.

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
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean69.931165068332741
Standard Deviation5.513.435.652354
CV%7.94.35.57.46.97.3

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=9) 101 85-116
EDTA Plasma (n=9) 97 88-107
Serum (n=11) 93 83-108
Linearity
To assess the linearity of the assay, five or more samples containing various concentrations of OPG in each matrix were diluted with Calibrator Diluent and then assayed.
Mouse Osteoprotegerin/TNFRSF11B Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Osteoprotegerin/TNFRSF11B
Osteoprotegerin (OPG), also known as OCIF and TNFRSF11B, is a secreted TNF superfamily receptor that regulates bone density. OPG is widely expressed and constitutively released as a homodimer by mesenchymal stem cells, fibroblasts, and endothelial cells. OPG functions as a decoy receptor for TRANCE/RANK L and TRAIL. OPG regulates the cell surface expression of TRANCE and its ability to stimulate osteoclastogenesis. Dysregulation of OPG/TRANCE interactions can lead to juvenile Paget’s disease, osteoporosis, bone loss, and vascular calcification.
    • Entrez Gene IDs
      4982 (Human); 18383 (Mouse);
    • Alternate Names
      OCIF; OCIFMGC29565; OPGtumor necrosis factor receptor superfamily member 11B; Osteoclastogenesis inhibitory factor; Osteoprotegerin; TNFRSF11B; TR1; tumor necrosis factor receptor superfamily, member 11b;
    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.

    32 Citations: Showing 1 - 10
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    Species
    Sample Type
    1. Role of glucocorticoid-induced leucine zipper (GILZ) in inflammatory bone loss
      Authors: N Yang, B Baban, CM Isales, XM Shi
      PLoS ONE, 2017;12(8):e0181133.
      Species: Mouse
      Sample Type: Serum
    2. Disruption of the Cx43/miR21 pathway leads to osteocyte apoptosis and increased osteoclastogenesis with aging
      Authors: HM Davis, R Pacheco-Co, EG Atkinson, LR Brun, AR Gortazar, J Harris, M Hiasa, SA Bolarinwa, T Yoneda, M Ivan, A Bruzzaniti, T Bellido, LI Plotkin
      Aging Cell, 2017;0(0):.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    3. Silicon Nitride: A Synthetic Mineral for Vertebrate Biology
      Sci Rep, 2016;6(0):31717.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Deletion of FoxO1, 3 and 4 in Osteoblast Progenitors Attenuates the Loss of Cancellous Bone Mass in a Mouse Model of Type 1 Diabetes
      J Bone Miner Res, 2016;0(0):.
      Species: Mouse
      Sample Type: Plasma
    5. 5-Azacytidine-induced protein 2 (AZI2) regulates bone mass by fine-tuning osteoclast survival.
      Authors: Maruyama K, Fukasaka M, Uematsu S, Takeuchi O, Kondo T, Saitoh T, Martino M, Akira S
      J Biol Chem, 2015;290(15):9377-86.
      Species: Mouse
      Sample Type: Serum
    6. Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-kappaB Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts.
      Authors: Kassem A, Henning P, Lundberg P, Souza P, Lindholm C, Lerner U
      J Biol Chem, 2015;290(33):20147-58.
      Species: Mouse
      Sample Type: Tissue Homogenates
    7. Effects of neuropeptides and mechanical loading on bone cell resorption in vitro.
      Authors: Yoo Y, Kwag J, Kim K, Kim C
      Int J Mol Sci, 2014;15(4):5874-83.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    8. Osteocytes produce interferon-beta as a negative regulator of osteoclastogenesis.
      Authors: Hayashida C, Ito J, Nakayachi M, Okayasu M, Ohyama Y, Hakeda Y, Sato T
      J Biol Chem, 2014;289(16):11545-55.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    9. Aconitum pseudo-laeve var. erectum inhibits receptor activator of nuclear factor kappa-B ligand-induced osteoclastogenesis via the c-Fos/nuclear factor of activated T-cells, cytoplasmic 1 signaling pathway and prevents lipopolysaccharide-induced bone loss in mice.
      Authors: Baek, Jong Min, Kim, Ju-Young, Cheon, Yoon-Hee, Park, Sun-Hyan, Ahn, Sung-Jun, Yoon, Kwon-Ha, Oh, Jaemin, Lee, Myeung S
      Molecules, 2014;19(8):11628-44.
      Species: Mouse
      Sample Type: Serum
    10. Bisphosphonates inhibit osteosarcoma-mediated osteolysis via attenuation of tumor expression of MCP-1 and RANKL.
      Authors: Ohba T, Cole H, Cates J, Slosky D, Haro H, Ando T, Schwartz H, Schoenecker J
      J Bone Miner Res, 2014;29(6):1431-45.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    11. Inhibition of bone remodeling in young mice by bisphosphonate displaces the plasma cell niche into the spleen.
      Authors: Teufel S, Grotsch B, Luther J, Derer A, Schinke T, Amling M, Schett G, Mielenz D, David J
      J Immunol, 2014;193(1):223-33.
      Species: Mouse
      Sample Type: Serum
    12. Resolvin E1 and chemokine-like receptor 1 mediate bone preservation.
      Authors: Gao L, Faibish D, Fredman G, Herrera B, Chiang N, Serhan C, Van Dyke T, Gyurko R
      J Immunol, 2013;190(2):689-94.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    13. Impaired bone formation and increased osteoclastogenesis in mice lacking chemokine (C-C motif) ligand 5 (Ccl5).
      Authors: Wintges K, Beil F, Albers J, Jeschke A, Schweizer M, Claass B, Tiegs G, Amling M, Schinke T
      J Bone Miner Res, 2013;28(10):2070-80.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    14. Strontium fructose 1,6-diphosphate prevents bone loss in a rat model of postmenopausal osteoporosis via the OPG/RANKL/RANK pathway.
      Authors: Ma B, Zhang Q, Wu D, Wang YL, Hu YY, Cheng YP, Yang ZD, Zheng YY, Ying HJ
      Acta Pharmacol. Sin., 2012;33(4):479-89.
      Species: Rat
      Sample Type: Serum
    15. Soluble rank ligand produced by myeloma cells causes generalised bone loss in multiple myeloma.
      Authors: Buckle, Clive He, De Leenheer, Evy, Lawson, Michelle, Yong, Kwee, Rabin, Neil, Perry, Mark, Vanderkerken, Karen, Croucher, Peter Ia
      PLoS ONE, 2012;7(8):e41127.
      Species: Mouse
      Sample Type: Serum
    16. The RANKL distal control region is required for the increase in RANKL expression, but not the bone loss, associated with hyperparathyroidism or lactation in adult mice.
      Authors: Onal M, Galli C, Fu Q, Xiong J, Weinstein RS, Manolagas SC, O'Brien CA
      Mol. Endocrinol., 2012;26(2):341-8.
      Species: Mouse
      Sample Type: Plasma
    17. Osteoprotection by semaphorin 3A.
      Authors: Hayashi M, Nakashima T, Taniguchi M, Kodama T, Kumanogoh A, Takayanagi H
      Nature, 2012;485(7396):69-74.
      Species: Mouse
      Sample Type: Serum
    18. Effects of relaxin and estrogens on bone remodeling markers, receptor activator of NF-kB ligand (RANKL) and osteoprotegerin (OPG), in rat adjuvant-induced arthritis.
      Authors: Ho TY, Santora K, Chen JC, Frankshun AL, Bagnell CA
      Bone, 2011;48(6):1346-53.
      Species: Rat
      Sample Type: Plasma
    19. Systemic effects of ulna loading in male rats during functional adaptation.
      Authors: Sample SJ, Collins RJ, Wilson AP, Racette MA, Behan M, Markel MD, Kalscheur VL, Hao Z, Muir P
      J. Bone Miner. Res., 2010;25(9):2016-28.
      Species: Rat
      Sample Type: Plasma
    20. Increased trabecular bone formation in mice lacking the growth factor midkine.
      Authors: Neunaber C, Catala-Lehnen P, Beil FT
      J. Bone Miner. Res., 2010;25(8):1724-35.
      Species: Mouse
      Sample Type: Serum
    21. Mecp2 deficiency decreases bone formation and reduces bone volume in a rodent model of Rett syndrome.
      Authors: O'Connor RD, Zayzafoon M, Farach-Carson MC, Schanen NC
      Bone, 2009;45(2):346-56.
      Species: Mouse
      Sample Type: Serum
    22. ADA-deficient SCID is associated with a specific microenvironment and bone phenotype characterized by RANKL/OPG imbalance and osteoblast insufficiency.
      Authors: Sauer AV, Mrak E, Hernandez RJ, Zacchi E, Cavani F, Casiraghi M, Grunebaum E, Roifman CM, Cervi MC, Ambrosi A, Carlucci F, Roncarolo MG, Villa A, Rubinacci A, Aiuti A
      Blood, 2009;114(15):3216-26.
      Species: Mouse
      Sample Type: Serum
    23. Beta-Arrestin2 regulates RANKL and ephrins gene expression in response to bone remodeling in mice.
      Authors: Pierroz DD, Rufo A, Bianchi EN, Glatt V, Capulli M, Rucci N, Cavat F, Rizzoli R, Teti A, Bouxsein ML, Ferrari SL
      J. Bone Miner. Res., 2009;24(5):775-84.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    24. Osteoprotegerin inhibits vascular calcification without affecting atherosclerosis in ldlr(-/-) mice.
      Authors: Morony S, Tintut Y, Zhang Z, Cattley RC, Van G, Dwyer D, Stolina M, Kostenuik PJ, Demer LL
      Circulation, 2008;117(3):411-20.
      Species: Mouse
      Sample Type: Plasma
    25. Myeloma-derived Dickkopf-1 disrupts Wnt-regulated osteoprotegerin and RANKL production by osteoblasts: a potential mechanism underlying osteolytic bone lesions in multiple myeloma.
      Authors: Qiang YW, Chen Y, Stephens O, Brown N, Chen B, Epstein J, Barlogie B, Shaughnessy JD
      Blood, 2008;112(1):196-207.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    26. Osteoprotegerin reduces the serum level of receptor activator of NF-kappaB ligand derived from osteoblasts.
      Authors: Nakamichi Y, Udagawa N, Kobayashi Y, Nakamura M, Yamamoto Y, Yamashita T, Mizoguchi T, Sato M, Mogi M, Penninger JM, Takahashi N
      J. Immunol., 2007;178(1):192-200.
      Species: Mouse
      Sample Type: Serum
    27. EGF-like ligands stimulate osteoclastogenesis by regulating expression of osteoclast regulatory factors by osteoblasts: implications for osteolytic bone metastases.
      Authors: Zhu J, Jia X, Xiao G, Kang Y, Partridge NC, Qin L
      J. Biol. Chem., 2007;282(37):26656-64.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    28. Activation of the p53 pathway down-regulates the osteoprotegerin expression and release by vascular endothelial cells.
      Authors: Secchiero P, Corallini F, Rimondi E, Chiaruttini C, di Iasio MG, Rustighi A, Del Sal G, Zauli G
      Blood, 2007;111(3):1287-94.
      Species: Mouse
      Sample Type: Serum
    29. B cells and T cells are critical for the preservation of bone homeostasis and attainment of peak bone mass in vivo.
      Authors: Li Y, Toraldo G, Li A, Yang X, Zhang H, Qian WP, Weitzmann MN
      Blood, 2007;109(9):3839-48.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    30. Osteoprotegerin in the inner ear may inhibit bone remodeling in the otic capsule.
      Authors: Zehnder AF, Kristiansen AG, Adams JC, Merchant SN, McKenna MJ
      Laryngoscope, 2005;115(1):172-7.
      Species: Mouse
      Sample Type: Serum
    31. JunB as a downstream mediator of PTHrP actions in cementoblasts.
      Authors: Berry JE, Ealba EL, Pettway GJ, Datta NS, Swanson EC, Somerman MJ, McCauley LK
      J. Bone Miner. Res., 2005;21(2):246-57.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    32. IL-17 promotes bone erosion in murine collagen-induced arthritis through loss of the receptor activator of NF-kappa B ligand/osteoprotegerin balance.
      Authors: Lubberts E, van den Bersselaar L, Oppers-Walgreen B, Schwarzenberger P, Coenen-de Roo CJ, Kolls JK, Joosten LA, van den Berg WB
      J. Immunol., 2003;170(5):2655-62.
      Species: Mouse
      Sample Type: Synovial Fluid
    Expand to show all 32 Citations
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    Summary

    Sample TestedSerum

    Other Experimental Details

    Other Experimental DetailsSerum samples had to be diluted 1:10 & 1:20 to be within range

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