Uncoated Human RANκL ( Receptor Activator of Nuclear Factor Kappa B Ligand ) ELISA Kit (E-UNEL-H0385)

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to RANκL . Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for RANκL and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain RANκL , biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of RANκL . You can calculate the concentration of RANκL in the samples by comparing the OD of the samples to the standard curve.

RANKL gene encodes a type II membrane protein of 316 amino acids with a predicted molecular mass of 35 kD. RANKL is cleaved to produce a soluble form with biological activity. The shedding of membrane-bound RANKL appears to be mediated by expression of MMP14 and ADAM10. Suppression of MMP14 in primary osteoblasts increases membrane-bound RANKL and promotes osteoclastogenesis in cocultures with macrophages. Therefore,RANKL shedding seems to be an important process that down-regulates local osteoclastogenesis. Alternatively,an increased production of RANKL by osteoblastic cells leads to osteoclast differentiation,activation,and survival,which results in increased bone resorption. Binding of RANKL to its receptor RANK activates TNF receptor-associated factor 6 (TRAF6),which is linked to downstream pathways including NF-κB,c-jun N-terminal kinase (JNK) or Src.