Osteoprotegerin Human ELISA

Research topic

Bone and cartilage metabolism

Features

• The total assay time is less than four hours.
• The kit measures total osteoprotegerin.
• Quality Controls are human serum based. No animal sera are used.

Storage/Shipping

Store the kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Summary

Osteoprotegerin (OPG) or osteoclastogenesis inhibitory factor (OCIF) is a secretory glycoprotein belonging to TNF receptor superfamily. OPG consists of 401 amino acid residues, it has a molecular weight of 60 kDa as a monomer and 120 kDa as a disulfide-linked dimer and is produced in different tissues, e.g. bone, skin, liver, stomach, intestine and lung. Osteoprotegerin inhibits the recruitment, proliferation and activation of osteoclasts. Osteoclast formation activity may be monitored principally by determination of concentration ratio of osteoprotegerin ligand (OPGL)/OPG. Alteration of this ratio may be the cause of bone loss in many imbalances in bone metabolism such as osteoporosis, osteopetrosis, hypercalcemia, metastatic osteolytic lesions and rheumatic bone degradation.

Assay format

Sandwich ELISA, Biotin-labelled antibody

Sample requirements

35 µl/well

Applications

Cell culture medium, Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum, Synovial fluid

Calibration Curve

Calibration range

1.5 – 60 pmol/l

Limit of detection

Analytical Limit of Detection is calculated from the real osteoprotegerin values in wells and is 0.13 pmol/l.

Limit of quantification

Assay Sensitivity takes the dilution of samples into consideration and is calculated according to the formula: Assay Sensitivity = Analytical Limit of Detection x sample dilution
= 0.13 pmol/l x 3 = 0.4 pmol/l.

Intra-assay (Within-Run, n=8)

CV = 4.5 %

Inter-assay (Run-to-Run, n=3)

CV = 5.5 %

Spiking Recovery

94.6 %

Dilution Linearity

103.2 %

Cross-Reactivity

Human

References to this product

• Clancy P, Oliver L, Jayalath R, Buttner P, Golledge J . Assessment of a serum assay for quantification of abdominal aortic calcification. Arterioscler Thromb Vasc Biol . Nov;26(11):2574-6 (2006)
• Maimoun L, Couret I, Mariano-Goulart D, Dupuy AM, Micallef JP, Peruchon E, Ohanna F, Cristol JP, Rossi M, Leroux JL. Changes in osteoprotegerin/RANKL system, bone mineral density, and bone biochemicals markers in patients with recent spinal cord injury. Calcif Tissue Int. 2005 Jun;76 (6):404-11
• Rogers A, Eastell R. Circulating osteoprotegerin and receptor activator for nuclear factor kappaB ligand: clinical utility in metabolic bone disease assessment. J Clin Endocrinol Metab. 2005 Nov;90 (11):6323-31
• Kanzaki H, Chiba M, Sato A, Miyagawa A, Arai K, Nukatsuka S, Mitani H. Cyclical tensile force on periodontal ligament cells inhibits osteoclastogenesis through OPG induction. J Dent Res. 2006 May;85 (5):457-62
• Avbersek-Luznik I, Balon BP, Rus I, Marc J. Increased bone resorption in HD patients: is it caused by elevated RANKL synthesis?. Nephrol Dial Transplant. 2005 Mar;20 (3):566-70
• Dai SM, Nishioka K, Yudoh K. Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha. Ann Rheum Dis. 2004 Nov;63 (11):1379-86
• Skladal P, Jilkova Z, Svoboda I, Kolar V. Investigation of osteoprotegerin interactions with ligands and antibodies using piezoelectric biosensors. Biosens Bioelectron. 2005 Apr 15;20 (10):2027-34
• Stejskal D, Zurek M, Bartek J, Jedelsky L, Ruzicka V. Osteoprotegerin and bone density. Biomed Pap Med Fac Univ Palack. 2001 Dec;145 (2):75-6
• Ozkaya O, Buyan N, Bideci A, Gonen S, Ortac E, Fidan K, Cinaz P, Soylemezoglu O. Osteoprotegerin and RANKL serum levels and their relationship with serum ghrelin in children with chronic renal failure and on dialysis. Nephron Clin Pract. 2007;105 (4):c153-8
• Avignon A, Sultan A, Piot C, Elaerts S, Cristol JP, Dupuy AM. Osteoprotegerin is associated with silent coronary artery disease in high-risk but asymptomatic type 2 diabetic patients. Diabetes Care. 2005 Sep;28 (9):2176-80
• Gogo PB Jr, Schneider DJ, Terrien EF, Sobel BE, Dauerman HL. Osteoprotegerin is not associated with angiographic coronary calcification. J Thromb Thrombolysis. 2006 Dec;22 (3):177-83
• Stejskal D, Bartek J, Pastorkova R, Ruzicka V, Oral I, Horalik D. Osteoprotegerin, RANK, RANKL. Biomed Pap Med Fac Univ Palack. 2001 Dec;145 (2):61-4
• Morena M, Terrier N, Jaussent I, Leray-Moragues H, Chalabi L, Rivory JP, Maurice F, Delcourt C, Cristol JP, Canaud B, Dupuy AM. Plasma osteoprotegerin is associated with mortality in hemodialysis patients. J Am Soc Nephrol. 2006 Jan;17 (1):262-70
• Golledge J, Jayalath R, Oliver L, Parr A, Schurgers L, Clancy P. Relationship between CT anthropometric measurements, adipokines and abdominal aortic calcification. Atherosclerosis. 2008 Mar;197 (1):428-34
• Garcia-Valdecasas-Campelo E, Gonzalez-Reimers E, Santolaria-Fernandez F, De la Vega-Prieto MJ, Milena-Abril A, Sanchez-Perez MJ, Martinez-Riera A, Gomez-Rodriguez Mde L. Serum osteoprotegerin and RANKL levels in chronic alcoholic liver disease. Alcohol Alcohol. 2006 May-Jun;41 (3):261-6
• Kim SM, Lee J, Ryu OH, Lee KW, Kim HY, Seo JA, Kim SG, Kim NH, Baik SH, Choi DS, Choi KM. Serum osteoprotegerin levels are associated with inflammation and pulse wave velocity. Clin Endocrinol (Oxf). 2005 Nov;63 (5):594-8
• Guldiken B, Guldiken S, Turgut B, Turgut N, Demir M, Celik Y, Arikan E, Tugrul A. Serum osteoprotegerin levels in patients with acute atherothrombotic stroke and lacunar infarct. Thromb Res. 2007;120 (4):511-6
• Kyrtsonis MC, Vassilakopoulos TP, Siakantaris MP, Kokoris SI, Gribabis DA, Dimopoulou MN, Angelopoulou MK, Pangalis GA. Serum syndecan-1, basic fibroblast growth factor and osteoprotegerin in myeloma patients at diagnosis and during the course of the disease. Eur J Haematol. 2004 Apr;72 (4):252-8

References to summary

• Ueland T. et all: Increased serum osteoprotegerin in disorders characterised by persistent immune activation or glucocorticoid excess – possible role in bone homeostasis. Europ. J. Endocrin. 145, 685–690 (2001)
• Feuerherm A. J. et all: Elevated levels of osteoprotegerin (OPG) and hepatocyte growth factor (HGF) in rheumatoid arthritis. Scand. J. Rheumatol. 30, 229–34 (2001)
• Yano K. et all: Immunological characterisation of Circulating osteoprotegerin/os­teoclastogene­sis inhibitory factor: Increased serum concentrations in postmenousal women with osteoporosis. J. of bone and mineral res. 4(14), 518– 527, (1999)
• Aubin J. E. and Bonnelye E.: Osteoprotegerin and its ligand: a new paradigm for regulation of osteoclastogenesis and bone resorption. Medscape Women Health 5 (2000)
• Hofbauer L. C.: Osteoprotegerin ligand and osteorotegerin: novel implications for osteoclast biology and bone metabolism. European Journal of Endocrinology 141, 195–210 (1999)
• Bucay N. et al.: Osteoprotegerin-deficient mice develop early onset osteoporosis and artificial calcification. Genes and Development 12, 1260–1268 (1998)
• Simonet W. S. et al.: Osteoprotegerin, a novel secreted protein involved in the regulation of bone density. Cell 89, 309–319 (1997)