本试剂盒应用双抗体夹心酶标免疫分析法测定标本中IL-27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-27抗原、生物素化的抗大鼠IL-27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-27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。
Intended use
This immunoassay kit allows for the specific measurement of rat IL-27 concentrations in cell culture supernates, serum, and plasma.
Introduction
Interleukin 27 (IL-27) is a non-covalently linked heterodimeric cytokine that is structurally related to IL-12. It is composed of an Epstein-Barr virus-induced molecule 3 (EBI3) subunit linked with the IL-27 p28 subunit. IL-27 is produced by activated antigen presenting cells including monocytes, endothelial cells, and dendritic cells. Its expression is induced in response to various inflammatory stimuli.IL-27 binds and signals through a heterodimeric receptor complex consisting of WSX-1 (TCCR) and gp130, both belonging to the cytokine receptor superfamily. WSX-1 is specific for IL-27 and is expressed on resting/naive CD4+ T cells, CD8+ T cells, NK cells, dendritic cells, monocytes, mast cells, and B cells. In contrast, gp130 is ubiquitously expressed by a variety of immune and non-immune cells and functions as a subunit of the receptor complexes for at least seven other cytokines. IL-27 has both pro- and anti-inflammatory properties. In response to infection, IL-27 induces monocytes and mast cells to secrete pro-inflammatory cytokines. It induces naive CD4+ T cells to proliferate and develop Th1 cell responses. IL-27 also promotes effector functions of NK cells and CD8+ T cells. As an anti-inflammatory immunomodulator, IL-27 has been found to have the ability to inhibit Th1 or Th2 responses and restrict the strength and duration of adaptive immune responses.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-27 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-27 present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for IL-27 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-27 bound in the initial step. The color development is stopped and the intensity of the color is measured.
