预期应用
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-17含量。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定标本中IL-17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-17抗原、生物素化的抗小鼠IL-17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-17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 mouse IL-17 concentrations in cell culture supernates, serum, and plasma.
Introduction
Interleukin 17 (IL-17; also known as IL-17A) is a 30 - 35 kDa variably glycosylated homodimeric protein that belongs to a unique family of cysteine-knot related proteins. Its sequence was originally isolated from an activated rodent hybridoma and termed CTLA-8 . It is synthesized as a 155 amino acid (aa) precursor that contains a 23 aa signal sequence and a 15 kDa, 132 aa mature segment. Although there are two intrachain disulfide bonds that create a ring reminiscent of those found in cysteine-knot proteins, the actual closed knot structure does not appear to form. IL-17 has one potential N-linked glycosylation site. Mature rat IL-17 is 61% and 60% aa identical to mouse and rat IL-17, respectively. It also shows limited aa sequence identity to other rat IL-17 family members. In particular, it shows 34%, 38%, 34% and 26%aa sequence identity to IL-17B, C, D, and E, respectively, and 49% aa sequence identity to IL-17F with which it is most homologous. While rodent and rat mature sequences show modest aa sequence identity, rat IL-17 is active on both mouse and rat cells. The cells principally known to produce IL-17 are the memory CD4+ T cells. In addition, CD8+ T cells as well as TCR+ CD4-CD8- T cells, neutrophils (PMNs) and eosinophils have also been reported to express mRNA transcripts for IL-17.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for IL-17 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-17 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for IL-17 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-17 bound in the initial step. The color
development is stopped and the intensity of the color is measured.
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