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血管性血友病因子裂解蛋白酶(ADAMTS13)也被称为ADAMTS13,是一种含锌的金属蛋白酶,可裂解von Willebrand factor(vWf),一种参与血液凝固的大蛋白。它被分泌到血液中并降解大型vWf多聚体,降低其活性。ADAMTS13基因映射到第九号染色体(9q34)。 从基因组上看,ADAMTS13与 ADAMTS家族的19个成员有许多共同的特性,所有这些成员都有一个蛋白酶结构域、一个相邻的分解蛋白结构域和一个或多个凝血酶结构域。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠金属肽酶含血小板反应蛋白13(ADAMTS13)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小鼠金属肽酶含血小板反应蛋白13(ADAMTS13)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 Mouse ADAMTS13. 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 Mouse ADAMTS13 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.