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Osteopontin(OPN),又称骨髓蛋白I(BSP-1或BNSP)、早期T淋巴细胞激活(ETA-1)、分泌型磷蛋白1(SPP1)、2ar和立克次体抵抗(Ric),由SPP1基因(分泌型磷蛋白1)编码的蛋白质。鼠类的同源物是Spp1。Osteopontin是一种SIBLING(糖蛋白),1986年首次在成骨细胞中发现。前缀osteo-表示该蛋白在骨中表达,尽管它也在其他组织中表达。后缀-pontin来源于拉丁语中的 "pons",意为桥梁,表示Osteopontin作为一种连接蛋白的作用。Osteopontin是一种细胞外结构蛋白,因此是骨骼的有机组成部分。 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桥素(OPN)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桥素(OPN)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 OPN. 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 OPN 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.