sRANKL (total) Human ELISA (Osteoprotegerin Ligand)
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
The RD193004200R Human sRANKL (total) ELISA is a sandwich enzyme immunoassay for the quantitative measurement of total sRANKL (free and bound sRANKL) in serum and plasma samples.
In the BioVendor Human sRANKL (total) ELISA, standards, quality controls and samples are incubated in microplate wells pre-coated with monoclonal anti-human sRANKL antibody. After 16-20 hours incubation and washing, biotin-labelled polyclonal anti-human sRANKL antibody is added and incubated with captured sRANKL for 60 minutes. After another washing, streptavidin-HRP conjugate is added. After 60 minutes incubation and the last washing step, the remaining conjugate is allowed to react with the substrate solution (TMB). The reaction is stopped by addition of acidic solution and absorbance of the resulting yellow product is measured spectrophotometrically at 450 nm. The absorbance is proportional to the concentration of sRANKL. A standard curve is constructed by plotting absorbance values against concentrations of standards, and concentrations of unknown samples are determined using this standard curve.
Intended use
Clinical Application
Test principle
Summary of protocol
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