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RAC-α丝氨酸/苏氨酸蛋白激酶由 AKT1 基因编码。该酶属于 AKT 亚家族的丝氨酸/苏氨酸激酶,含有 SH2(Src 同源性 2-like)蛋白结构域。它通常被称为 PKB,或同时被称为 "Akt/PKB"。丝氨酸-苏氨酸蛋白激酶 AKT1 在血清干燥的原生和永生成纤维细胞中没有催化作用。AKT1 和相关的 AKT2 被血小板衍生的生长因子激活。缺乏 Akt1 的小鼠显示身体质量减少 25%,表明 Akt1 对于传递促进生长的信号至关重要,很可能是通过 IGF1 受体。缺少Akt1 的小鼠对癌症也有抵抗力。它们在由大 T 抗原或 Neu 肿瘤基因启动的肿瘤生长中经历了相当大的延迟。 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AKT1蛋白(AKT1)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AKT1蛋白(AKT1)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 AKT1. 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 AKT1 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.