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Tau 蛋白是一种高度可溶性的微管相关蛋白(MAP)。与非神经元细胞相比, 这些蛋白主要存在于神经元中。Tau 是一种磷酸化蛋白,在最长的 tau 异构 体上有 79 个潜在的丝氨酸(Ser)和苏氨酸(Thr)磷酸化点。异常高磷酸 化的微管相关蛋白 Tau(p-Tau)通过与管蛋白/微管竞争与正常 Tau、MAP1 和 MAP2 结合,抑制微管的组装并破坏微管。p-Tau 可被蛋白磷酸酶 PP-2B、 PP-2A 和 PP-1 去磷酸化,这些都存在于神经元。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠磷酸化Tau蛋白(p-Tau)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小鼠磷酸化Tau蛋白(p-Tau)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 Mouse p-Tau. 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 Mouse p-Tau 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.