Description
Background: Receptor activator of nuclear factor kappa-B ligand (RANKL), also known as tumor necrosis factor ligand superfamily member 11 (TNFSF11), is a protein that in humans is encoded by the TNFSF11 gene. This gene encodes a member of the tumor necrosis factor (TNF) cytokine family which is a ligand for osteoprotegerin and functions as a key factor for osteoclast differentiation and activation. This gene is mapped to chromosome13q14.11. Targeted disruption of the related gene in mice led to severe osteopetrosis and a lack of osteoclasts. The deficient mice exhibited defects in early differentiation of T and B lymphocytes, and failed to form lobulo-alveolar mammary structures during pregnancy. This gene may play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy.
Principle of the Assay: The Picokine™ Human TNFSF11 Pre-Coated ELISA (Enzyme-Linked Immunosorbent Assay) kit is a solid-phase immunoassay specially designed to measure Human TNFSF11 with a 96-well strip plate that is pre-coated with antibody specific for TNFSF11. The detection antibody is a biotinylated antibody specific for TNFSF11. The capture antibody is polyclonal antibody from goat and the detection antibody is polyclonal antibody from goat. The kit includes Human TNFSF11 protein as standards. To measure Human TNFSF11, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS or TBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS or TBS buffer and add TMB. TMB is an HRP substrate and will be catalyzed to produce a blue color product, which changes into yellow after adding the acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to Human TNFSF11 in the sample. Read the absorbance of the yellow product in each well using a plate reader, and benchmark the sample wells' readings against the standard curve to determine the concentration of Human TNFSF11 in the sample