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α平滑肌肌动蛋白(α-SMA)通常称为平滑肌肌动蛋白α2(ACTa2),常被用 作肌成纤维细胞形成的标志物。ACTa2 是 6 种不同的肌动蛋白异构体之一, 是参与平滑肌的收缩装置。它与所有的肌动蛋白一样是高度保守的,几乎在 所有的哺乳动物中都能找到。它由位于 10q22-q24 的 ACTA2 基因编码。该 基因的突变导致各种血管疾病,如胸主动脉疾病、冠状动脉疾病、中风、 Moyamoya 病和多系统平滑肌功能障碍综合症。 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α平滑肌肌动蛋白(α-SMA)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α平滑肌肌动蛋白(α-SMA)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 α-SMA. 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 α-SMA 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.