Receptor Activator of Nuclear Factor k 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 383aa in its intracellular domain(residues 234-616), which contain three putative TRAF-binding domains(termed I, II, and III). 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.
Sample Type
Cell culture supernates and serum.
Intended Use
For quantitative detection of rat RANK in cell culture supernates and serum.
Specificity
Natural and recombinant rat RANK
Test Principle
This rat RANK BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for RANK has been precoated onto 96-well plates. Standards(NSO, Q30-P213) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for RANK 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 RANK amount of sample captured in plate.
Kit Components
96-well plate precoated with rat RANK antibody, 1 Lyophilized recombinant rat RANK standard, 10ng/tube×2 Biotinylated rat RANK antibody, 130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100) Sample diluent buffer, 30ml Antibody diluent buffer, 12ml ABC diluent buffer, 12ml TMB color developing agent, 10ml TMB stop solution, 10ml
Storage and Stability
Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
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