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甲硫氨酰氨肽酶 2 是由 METAP2 基因编码的酶。它是一种胞质溶胶金属酶,其催化从新生蛋白质中水解去除 N 端甲硫氨酸残基。肽-蛋氨酸\rightleftharpoons 肽+蛋氨酸 MetAP2 存在于所有生物中,由于其在组织修复和蛋白质降解中的关键作用而尤为重要。与 METAP2 相关的疾病包括小孢子菌病和肝脏横纹肌肉瘤。其相关途径包括视觉光传导和 Arg/N 端规则途径(真核生物)。与该基因相关的基因本体论注释包括 RNA 结合和氨基肽酶活性。该基因的一个重要类似物是 PA2G4。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人甲硫氨酰氨肽酶2(METAP2)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甲硫氨酰氨肽酶2(METAP2)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 METAP2. 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 METAP2 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.