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骨保护素(OPG),也被称为破骨细胞生成抑制因子(OCIF)或肿瘤坏死因子受体超家族成员11B(TNFRSF11B),是由TNFRSF11B基因编码的肿瘤坏死因子(TNF)受体超家族的一个细胞因子受体。OPG是一种可溶性的糖蛋白,可以是60kDa的单体,也可以是120kDa的二聚体,由二硫键连接。它由380个氨基酸组成,形成7个功能域。1-4域是富含半胱氨酸的N端域,在结合过程中与RANKL相互作用。5-6域是死亡域,有助于OPG的二聚化。7域是C端肝素结合域,以一个半胱氨酸(Cys-400)结束,也在OPG的二聚化中发挥重要作用。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有大鼠骨保护素(OPG)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大鼠骨保护素(OPG)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 Rat OPG. 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 Rat OPG 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.