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软骨寡聚基质蛋白(COMP),也被称为血栓软骨素-5,是一种主要存在于软骨中的细胞外基质(ECM)蛋白,是由COMP基因编码的。该基因编码的蛋白是一种非胶原性细胞外基质(ECM)蛋白。它由五个相同的糖蛋白亚单位组成,每个亚单位都有EGF样和钙结合(血栓软骨素样)结构域。低聚物是由五股螺旋线圈和二硫键形成的。与其他ECM蛋白如胶原蛋白的结合似乎取决于二价阳离子。突变可导致骨软骨发育不良的假性软骨发育不良(PSACH)和多发性骺线发育不良(MED)。COMP是软骨周转的一个标志。它在纤维化疤痕和系统性硬化症中大量存在,而且它似乎在血管壁重塑中起作用。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人软骨寡聚基质蛋白(COMP)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软骨寡聚基质蛋白(COMP)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 COMP. 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 COMP 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.