sRANKL (total) Human ELISA (Osteoprotegerin Ligand)

Summary

sRANKL, receptor activator of nuclear factor (NF)-κB ligand (also: osteoprotegerin ligand, OPGL), is a part of the TNF superfamily with high similarity to other members of that protein species. (SwissProt Nr. O14788).

Three isoforms are produced by alternate splicing, two type II membrane proteins (ISOFORM 1, 317 AA, and ISOFORM 3, 270 AA), and a secreted molecule (ISOFORM 2, 244 AA). ISOFORM 1 is identical to previously reported RANKL and possesses intracellular, transmembrane, and extracellular domains; ISOFORM 2 does not have the intracellular and transmembrane domains, and ISOFORM 3 does not have the intracellular domain. A soluble form arises by proteolytic processing from membrane isoforms.

Although all forms are bioactive, the membrane-bound proteins seem to be the homeostatic forms, while the production of soluble RANKL signals pathological conditions.

RANKL, RANK, and osteoprotegerin (OPG) have been identified as the key molecular regulation system for bone remodelling. RANKL is the main stimulatory factor for the formation of mature osteoclasts and is essential for their survival. Therefore, an increase in RANKL expression leads to bone resorption and bone loss. RANKL is produced by osteoblastic lineage cells and activated T lymphocytes. It activates its specific receptor RANK, which is located on osteoclasts and dendritic cells. The effects of RANKL are counteracted by OPG, which is secreted by various tissues and acts as an endogenous soluble receptor antagonist.

Imbalances of the RANKL/OPG system have been related to the pathogenesis of Paget’s disease, benign and malignant bone tumors, postmenopausal osteoporosis, rheumatoid arthritis, bone metastases and hypercalcemia. Several studies using animal models have shown that restoring the RANKL/OPG balance (e.g. by administering OPG) reduces the severity of these disorders.

Indication

• Postmenopausal and senile osteoporosis
• Diseases with locally increased bone resorption activity
• Paget´s disease
• Periodontal disease
• Cardiovascular disease, arterial calcification
• Inflammatory diseases
• Immunological disorders
• Arthritis
• Oncology

Features

• It is intended for research use only
• The total assay time is about 20 hours
• The kit measures total sRANKL in serum and plasma (EDTA, citrate, heparin)
• Assay format is 96 wells
• Quality Controls are native protein based
• Standard is serum protein based
• Components of the kit are provided ready to use, concentrated or lyophilized

Research topic

Bone and cartilage metabolism

Assay format

Sandwich ELISA, Biotin-labelled antibody

Applications

Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum

Sample requirements

5 µl/well

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).

Calibration Curve

Calibration range

0.5 – 32 pmol/l

Limit of detection

0.4 pmol/l

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

CV = 8.70 %

Inter-assay (Run-to-Run, n=4)

CV = 9.19 %

Spiking Recovery

102.7 %

Dilution Linearity

100.4 %

Cross-Reactivity

References to this product

• Adamopoulos IE, Chao CC, Geissler R, Laface D, Blumenschein W, Iwakura Y, McClanahan T, Bowman EP. Interleukin-17A upregulates receptor activator of NF-kappaB on osteoclast precursors. Arthritis Res Ther. 2010;12 (1):R29
• Barbagallo I, Vanella A, Peterson SJ, Kim DH, Tibullo D, Giallongo C, Vanella L, Parrinello N, Palumbo GA, Di Raimondo F, Abraham NG, Asprinio D. Overexpression of heme oxygenase-1 increases human osteoblast stem cell differentiation. J Bone Miner Metab. 2010 May;28 (3):276-88
• Chandran V, Cook RJ, Edwin J, Shen H, Pellett FJ, Shanmugarajah S, Rosen CF, Gladman DD. Soluble biomarkers differentiate patients with psoriatic arthritis from those with psoriasis without arthritis. Rheumatology (Oxford). 2010 Jul;49 (7):1399-405
• Chen CH, Chen HA, Liao HT, Liu CH, Tsai CY, Chou CT. Soluble receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin in ankylosing spondylitis: OPG is associated with poor physical mobility and reflects systemic inflammation. Clin Rheumatol. 2010 Oct;29 (10):1155-61
• Christoforidis A, Economou M, Farmaki E, Tzimouli V, Gombakis N, Athanassiou-Metaxa M. Increased osteoclastic activity as shown by increased sRANK-L/OPG ratio in boys with hemophilia. Ann Hematol. 2010 Aug;89 (8):837-8
• Christoforidis A, Economou M, Papadopoulou E, Kazantzidou E, Farmaki E, Tzimouli V, Tsatra I, Gompakis N, Athanassiou-Metaxa M. Comparative study of dual energy X-ray absorptiometry and quantitative ultrasonography with the use of biochemical markers of bone turnover in boys with haemophilia. Haemophilia. 2011 Jan;17 (1):e217-22
• D'Amelio P, Cristofaro MA, D'Amico L, Veneziano L, Roato I, Sassi F, Bisignano G, Saracco M, Pellerito R, Patane S, Ferracini R, Pescarmona GP, Isaia GC. Iloprost modulates the immune response in systemic sclerosis. BMC Immunol. 2010;11:62
• Ford ML, Smith ER, Tomlinson LA, Chatterjee PK, Rajkumar C, Holt SG. FGF-23 and osteoprotegerin are independently associated with myocardial damage in chronic kidney disease stages 3 and 4. Another link between chronic kidney disease-mineral bone disorder and the heart. Nephrol Dial Transplant. 2011 Jul 12;
• 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 . Apr;72(4):252-8 (2004)
• Mohamed JA, DuPont HL, Jiang ZD, Flores J, Carlin LG, Belkind-Gerson J, Martinez-Sandoval FG, Guo D, White AC Jr, Okhuysen PC. A single-nucleotide polymorphism in the gene encoding osteoprotegerin, an anti-inflammatory protein produced in response to infection with diarrheagenic Escherichia coli, is associated with an increased risk of nonsecretory bacterial diarrhea in North American travelers to Mexico. J Infect Dis. 2009 Feb 15;199 (4):477-85
• Morena M, Bosc JY, Jaussent I, Dupuy AM, Terrier N, Leray-Moragues H, Flavier JL, Maurice F, Delcourt C, Cristol JP, Canaud B. The role of mineral metabolism and inflammation on dialysis vascular access failure. J Vasc Access. 2006 Apr-Jun;7 (2):77-82
• Nezami N, Safa J, Eftekhar-Sadat AT, Salari B, Ghorashi S, Sakhaee K, Khosraviani K. Lovastatin raises serum osteoprotegerin level in people with type 2 diabetic nephropathy. Clin Biochem. 2010 Nov;43 (16-17):1294-9
• Ozcaka O, Nalbantsoy A, Kose T, Buduneli N. Plasma osteoprotegerin levels are decreased in smoker chronic periodontitis patients. Aust Dent J. 2010 Dec;55 (4):405-10
• 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
• Ren W, Blasier R, Peng X, Shi T, Wooley PH, Markel D. Effect of oral erythromycin therapy in patients with aseptic loosening of joint prostheses. Bone. 2009 Apr;44 (4):671-7
• Tibullo D, Giallongo C, La Cava P, Berretta S, Stagno F, Chiarenza A, Conticello C, Palumbo GA, Di Raimondo F. Effects of imatinib mesylate in osteoblastogenesis. Exp Hematol. 2009 Apr;37 (4):461-8
• Zdzisinska B, Bojarska-Junak A, Walter-Croneck A, Kandefer-Szerszen M. Dysregulation of the receptor activator of NF-kappaB ligand and osteoprotegerin production influence the apoptosis of multiple myeloma patients' bone marrow stromal cells co-cultured with myeloma cells. Arch Immunol Ther Exp (Warsz). 2010 Apr;58 (2):153-63

References to summary

• Hofbauer L.C et al. Effect of oral contraceptives on circulatingos­teoprotegerin and soluble RANK ligand serum levels in healthy young women. Clin Endocrinol (2004) 60: 214–219
• Ito S. et al. Crystal structure of the extracellular domain of mouse ligand at2.2-A resolution. J Biol Chem (2002) 22: 6631–6636.
• Lam J. et al. Crystal structure of the TRANCE/RANKL cytokine reveals determinants of receptorligand specifity. J Clin Invest (2001) 108: 971–979.
• Lacey D.L, et al., Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell (1998), 93:165–176.
• Kong Y.Y. et al., OPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis. Nature (1999), 397: 315–323.
• Hsu H. et al., Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci (1999), 96:3540–3545.
• Josien R. et al, TRANCE, a tumor necrosis factor family member, enhances the longevity and adjuvant properties of dendritic cells in vivo. J Exp Med (2000), 191: 495–502.
• Fuller K. et al., TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. J Exp Med (1998), 188: 997–1001.
• Nakashima T. et. al., Protein expression and functional difference of membrane-bound and soluble receptor activator of NF-kappaB ligand: modulation of the expression by osteotropic factors and cytokines. Biochem Biophys Res Commun (2000), 275(3):768–775.
• Kong Y.Y. et al., Activated T cells regulate bone loss and joint destruction in adjuvant arthritis through osteoprotegerin ligand. Nature (1999), 402: 304–309.
• Hofbauer L.C. & A.E. Heufelder, Role of receptor activator of nuclear factor-KB ligand and osteoprotegerin in bone cell biology. J Mol Med (2001), 79: 243–253.
• Hofbauer L.C. & A.E. Heufelder, The Role of Osteoprotegerin and Receptor Activator of Nuclear Factor κB Ligand in the Pathogenesis and Treatment of Rheumatoid Arthritis. Arthritis & Rheumatism (2001), 44:253–259.
• Hofbauer LC, et al. The role of receptor activator of nuclear factor-kappaB ligand and osteoprotegerin in the pathogenesis and treatment of metabolic bone diseases. J Clin Endocrinol Metab (2000), 85: 2355–2363.
• Teitelbaum S.L., Bone resorption by osteoclasts. Science (2000), 289: 1504–1508.

By research topic:

• Bone and cartilage metabolism

By molecule:

• RANKL

Registration form