Interleukin 23 (IL-23) is a heterodimeric cytokine composed of two disulfide-linked subunits,a p19 subunit that is unique to IL-23,and a p40 subunit that is shared with IL-12. The p19 subunit has homology to the p35 subunit of IL-12,as well as to other single chain cytokines such as IL-6 and IL-11. The p40 subunit is homologous to the extracellular domains of the hematopoietic cytokine receptors. Mouse p19 cDNA encodes a 196aa residue precursor protein with a putative 19aa signal peptide and 177aa mature protein. Human and mouse p19 share 70%aa sequence identity. Although p19 is expressed by activated macrophages,dendritic cells,T cells,and endothelial cells,only activated macrophages and dendritic cells express p40 concurrently to produce IL-23. The functional IL-23 receptor complex consists of two receptor subunits,the IL-12 receptor beta 1 subunit (IL-12 R beta 1) and the IL-23-specific receptor subunit (IL-23 R). IL-23 has biological activities that are similar to,but distinct from IL-12. Both IL-12 and IL-23 induce proliferation and IFN-gamma production by human T cells. While IL-12 acts on both naive and memory human T cells,the effects of IL-23 is restricted to memory T cells. In mouse,IL-23 but not IL-12,has also been shown to induce memory T cells to secret IL-17,a potent proinflammatory cytokine. IL-12 and IL-23 can induce IL-12 production from mouse splenic DC of both the CD8- and CD8+ subtypes,however only IL-23 can act directly on CD8+ DC to mediate immunogenic presentation of poorly immunogenic tumor/self peptide.Sample Type:Cell culture supernatants,serum,plasma,and tissueIntended Use:The Human IL-23 BioAssay ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human IL-23 concentrations in cell culture supernatants,serum,plasma,and tissue.Sensitivity:15pg/mlRange:31.25-2000pg/mlSpecificity:Recognizes human IL-23.Sample Volume:100ul/wellTest Principle:This assay employs the Sandwich immunoassay technique. An anti-h IL-23 monoclonal coating antibody is adsorbed onto microwells. IL-23 present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h IL-23 antibody is added and binds to IL-23 captured by the first antibody. Following incubation unbound Biotinylated anti-h IL-23 antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h IL-23 antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of IL-23 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-23 standard dilutions and IL-23 sample concentration determined.Kit Components:Aluminium pouches with a Microwell Plate: 8x12Standard: 2xVialBiotinylated antibody: 2xVialStreptavidin-HRP: 2xVialAssay Buffer (25ml/Vial): 2xVialWash Buffer Concentrate 20x (30ml/Vial): 1xVialTMB Substrate Solution: 1x12mlStop Solution (12ml/Vial): 1xVialPlate Covers- Adhesive strips: 3xstripsInstruction: 1xStorage 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.