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巨噬细胞移动抑制因子(MIF)又称糖基化抑制因子(GIF)、L-多巴色异构酶或苯丙酮酸同构酶,是一种由MIF基因编码的蛋白质。它是先天性免疫的一个重要调节因子。 MIF蛋白超家族还包括第二个具有功能相关特性的成员,即D-多巴色异构酶(D-DT)。CD74是MIF的表面受体。细菌抗原刺激白细胞释放MIF进入血流。循环中的MIF与其他免疫细胞上的CD74结合,引发急性免疫反应。因此,MIF被归类为一种炎症性细胞因子。此外,糖皮质激素也刺激白细胞释放MIF,因此MIF部分地抵消了糖皮质激素对免疫系统的抑制作用。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人巨噬细胞移动抑制因子(MIF)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巨噬细胞移动抑制因子(MIF)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 MIF. 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 MIF 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.