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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
纤连蛋白(FN)是细胞外基质的一种高分子量(约500-600 kDa)糖蛋白,与称为整合素的跨膜受体蛋白结合。纤连蛋白还与其他细胞外基质蛋白结合,如胶原蛋白、纤维蛋白和硫酸肝素蛋白多糖。纤连蛋白以蛋白二聚体的形式存在,由两条几乎相同的多肽链组成,通过一对C端二硫键连接。每个纤连蛋白亚单位的氨基酸分子量为~230-~275 kDa,包含三种类型的模块:I型、II型和III型。所有这三种模块都是由两个反平行的β-片组成,形成一个Beta-三明治;然而,I型和II型是由链内二硫键稳定的,而III型模块不包含任何二硫键。III型模块中没有二硫键,这使得它们在受力时可以部分展开。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人纤连蛋白(FN)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纤连蛋白(FN)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 FN. 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 FN 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.