Mouse Osteoprotegerin/TNFRSF11B Quantikine ELISA Kit

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Mouse Osteoprotegerin/TNFRSF11B Standard Curve
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Mouse Osteoprotegerin/TNFRSF11B 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 (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
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 69.9 311 650 68 332 741
Standard Deviation 5.5 13.4 35.6 5 23 54
CV% 7.9 4.3 5.5 7.4 6.9 7.3

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 ELISA Linearity

Product Datasheets

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
⚠ 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 Osteoprotegerin/TNFRSF11B 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.

35 Citations: Showing 1 - 10
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  1. Soluble RANKL contributes to osteoclast formation in adult mice but not ovariectomy-induced bone loss
    Authors: J Xiong, K Cawley, M Piemontese, Y Fujiwara, H Zhao, JJ Goellner, CA O'Brien
    Nat Commun, 2018;9(1):2909.
    Species: Mouse
    Sample Types: Plasma
  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 Types: Cell Culture Supernates
  3. Muramyl Dipeptide, a Shared Structural Motif of Peptidoglycans, Is a Novel Inducer of Bone Formation through Induction of Runx2
    Authors: OJ Park, J Kim, J Yang, CH Yun, SH Han
    J. Bone Miner. Res, 2017;0(0):.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  4. Hypoxia-Inducible Factor 1-alpha does not regulate osteoclastogenesis but enhances bone resorption activity via prolyl-4-hydroxylase 2
    Authors: PA Hulley, T Bishop, A Vernet, JE Schneider, JR Edwards, NA Athanasou, HJ Knowles
    J. Pathol., 2017;0(0):.
    Species: Mouse
    Sample Types: Serum
  5. 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 Types: Serum
  6. Silicon Nitride: A Synthetic Mineral for Vertebrate Biology
    Sci Rep, 2016;6(0):31717.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. 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 Types: Plasma
  8. 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 Types: Serum
  9. 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 Types: Tissue Homogenates
  10. 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 Types: Cell Culture Supernates
  11. 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 Types: Cell Culture Supernates
  12. 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 Types: Serum
  13. 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 Types: Cell Culture Supernates
  14. 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 Types: Serum
  15. 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 Types: Cell Culture Supernates
  16. 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 Types: Cell Culture Supernates
  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 Types: Serum
  18. 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 Types: Serum
  19. 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 Types: Serum
  20. 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 Types: Plasma
  21. 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 Types: Plasma
  22. 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 Types: Plasma
  23. 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 Types: Serum
  24. 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 Types: Serum
  25. 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 Types: Serum
  26. 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 Types: Cell Culture Supernates
  27. 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 Types: Cell Culture Supernates
  28. 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 Types: Plasma
  29. 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 Types: Serum
  30. 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 Types: Serum
  31. 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 Types: Cell Culture Supernates
  32. 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 Types: Cell Culture Supernates
  33. 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 Types: Serum
  34. 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 Types: Cell Culture Supernates
  35. 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 Types: Synovial Fluid

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Mouse Osteoprotegerin/TNFRSF11B Quantikine ELISA Kit
By Anonymous on 08/10/2017
Sample Tested: Serum

Serum samples had to be diluted 1:10 & 1:20 to be within range