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. The rat gene is mapped to 15q11. 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. Sample Type: Cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Rat TNFSF11/RANKL. Sensitivity: <10pg/ml Range: 15.6pg/ml-1000pg/ml Specificity: Recognizes natural and recombinant Rat TNFSF11/RANKL. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Rat TNFSF11/RANKL BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune- sorbent assay technology. A specific for TNFSF11/RANKL has been precoated onto 96-well plates. Standards(Expression system for standard: E.coli, Immunogen sequence: F141-D318) and test samples are added to the wells, a biotinylated detection specific for TNFSF11/RANKL is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Rat TNFSF11/RANKL amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Rat Tnfsf11 antibody, 1x plate Lyophilized recombinant Rat Tnfsf11 standard, 2x10ng/tube Biotinylated anti-Rat Tnfsf11 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
