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EK000130-EK0829: Human RANK ELISA Kit
Range 62.5pg/ml-4000pg/ml
Sensitivity < 2 pg/ml
Specificity: No detectable cross-reactivity with any other cytokine.
Application: For quantitative detection of human RANK in tissue lysates or cell culture supernates, and should be confirmed for serum and plasma.
Expiration: Four months at 4°C and eight months at -20°C.
Background
Receptor Activator of Nuclear Factor B (RANK), also known as TRANCE Receptor, is a type I membrane protein that is expressed on the surface of osteoclasts and is involved in their activation upon ligand binding. RANK is a recently described TNF receptor family member, and its ligand, RANKL, promote survival of dendritic cells and differentiation of osteoclasts. RANK contains 383 amino acids in its intracellular domain (residues 234-616), which contain three putative TRAF-binding domains (termed I, II, and III). Darnay BG et al demonstrated that RANK interacts with various TRAFs through distinct motifs and activates NF-kappaB via a novel TRAF6 interaction motif, which then activates NIK, thus leading to NF-kappaB activation, whereas RANK most likely activates JNK through a TRAF2-interacting region in RANK. The standard in this kit is recombinant human RANK with the sequence of Q29-G213 aa. It is a dipolymer which compose
of two chains, and the molecular weight of each is 48kda.
Principle
RANK ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Human RANK specific-specific polyclonal antibodies were precoated onto 96-well plates. The human specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells 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 human RANK amount of sample captured in plate.
Reference
1. Darnay, B G; Haridas V, Ni J, Moore P A, Aggarwal B B (Aug. 1998). "Characterization of the intracellular domain of receptor activator of NF-kappaB (RANK). Interaction with tumor necrosis factor receptor-associated factors and activation of NF-kappab and c-Jun N-terminal kinase". J. Biol. Chem. (UNITED STATES) 273 (32): 20551?.
2. Darnay, B G; Ni J, Moore P A, Aggarwal B B (Mar. 1999). "Activation of NF-kappaB by RANK requires tumor necrosis factor receptor-associated factor (TRAF) 6 and NF-kappaB-inducing kinase. Identification of a novel TRAF6 interaction motif". J. Biol. Chem. (UNITED STATES) 274 (12): 7724?1.
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