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五聚素3(PTX3)也被称为TNF诱导基因14蛋白(TSG-14),由PTX3基因编码。PTX3是五联蛋白超家族的一个成员,这个超家族的特点是环状多聚体结构,它由几种细胞类型迅速产生和释放,特别是由单核吞噬细胞、树突状细胞(DCs)、成纤维细胞和内皮细胞对初级炎症信号的反应。PTX3与补体成分C1q、细胞外基质成分TNF-α诱导蛋白6和选定的微生物(包括烟曲霉和铜绿假单胞菌)有高度亲和力的结合。PTX3激活补体激活的经典途径,促进巨噬细胞和DCs识别病原体。它定位于3号染色体的合成区(q24-28),高度保守,有82%相同的氨基酸和92%保守的氨基酸。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人五聚素3(PTX3)捕获抗体的微孔中,依次加入样本、标准品、生物素标记的检测抗体,HRP酶结合物,中间经过温育和洗涤,用底物TMB显色,TMB在过氧化物酶(HRP)的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样本中的人五聚素3(PTX3)呈正相关。用酶标仪在450nm 波长下测定吸光度(OD值),计算样本浓度。
This kit employs the sandwich enzyme-linked immunosorbent assay (ELISA) principle. Microplate wells are pre-coated with a capture antibody specific to Human PTX3. Following this, the sample, standards, a biotin-conjugated detection antibody, and an HRP (horseradish peroxidase)-labeled conjugate are added sequentially to the wells. The procedure involves incubation and wash steps between each addition. For detection, TMB substrate is used. TMB yields a blue color in the presence of the peroxidase activity of HRP. The reaction is then stopped with an acid solution, which converts the blue color to a final yellow color. The intensity of the yellow color is directly proportional to the concentration of Human PTX3 present in the sample. The optical density (OD) is measured at a wavelength of 450 nm using a microplate reader. Finally, the concentration of the target analyte in the sample is calculated based on the standard curve generated from the standards.