本试剂盒应用双抗体夹心酶标免疫分析法测定标本中白细胞介素-10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白细胞介素-10抗原、生物素化的抗猪白细胞介素-10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白细胞介素-10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 porcine IL-10 concentrations in cell culture supernates, serum, and plasma.
Introduction
Interleukin 10, also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 cytokine family. This family currently comprises IL-10, IL-19, IL-20, IL-22, IL-24 and IL-26. All known IL-10 family members are secreted -helical proteins. IL-10 is a secreted, possibly glycosylated, polypeptide with a molecular weight of 18 kDa. Based on human studies, IL-10 is likely to circulate as a nondisulfide-linked homodimer. IL-10 is synthesized as a 181 amino acid (aa) precursor with a 19 aa signal sequence and a 162 aa mature form. The mature segment has one potential N-linked glycosylation site plus four cysteines which likely form two intra-chain disulfide bridges. Mature rat IL-10 shows 76%, 73%, 72%, 80%, 86%, 85% and 76% aa identity to guinea pig, mouse, human, feline and equine IL-10, respectively. Upon activation, mammalian cells known to secrete IL-10 include NK cells, cytotoxic CD8+ T cells secreting Th2-like cytokines, CD4+CD45RA- (memory) Th1 and Th2 cells, macrophages, monocytes, CD5+ and CD5- B cells, dendritic cells, hepatic stellate (Ito) cells, keratinocytes, melanoma cells, mast cells, placental cytotrophoblasts, and fetal erythroblasts.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for IL-10 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-10 present is bound by the immobilized antibody. An
enzyme-linked monoclonal antibody specific for IL-10 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-10 bound in the initial step. The color
development is stopped and the intensity of the color is measured.
