ELISA法定量测定小鼠血清、血浆、细胞培养物上清或其它相关液体中IL-23含量。

 

    本试剂盒应用双抗体夹心酶标免疫分析法测定标本中IL-23水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入IL-23抗原、生物素化的抗大鼠IL-23抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的IL-23呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。

 

    This immunoassay kit allows for the specific measurement of mouse IL-23 concentrations in cell culture supernates, serum, and plasma.

 

    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. Rat p19 cDNA encodes a 189 amino acid residue (aa) precursor protein with a putative 19 aa signal peptide and 170 aa mature protein. Rat 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 Rb1) 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-γ production by rat T cells. While IL-12 acts on both naïve and memory rat 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.

 

    This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-23 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-23 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for IL-23 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-23 bound in the initial step. The color development is stopped and the intensity of the color is measured.