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磷脂酰乙醇胺-N-甲基转移酶(PEMT)是一种转移酶(EC 2.1.1.17),在肝脏中将磷脂酰乙醇胺(PE)转换为磷脂酰胆碱(PC)。在 CDP-胆碱途径中, 通过饮食或含胆碱的脂质代谢获得的胆碱被转化为 PC,约占肝脏中 PC 生物合成的 70%,而 PEMT 途径已被证明在饥饿时提供 PC 方面发挥了关键的进化作用。此外,通过 PEMT 制造的 PC 发挥了广泛的生理作用,用于胆碱合成、肝细胞膜结构、胆汁分泌和极低密度脂蛋白(VLDL)的分泌。 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脂酰乙醇胺-N-甲基转移酶(PEMT)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脂酰乙醇胺-N-甲基转移酶(PEMT)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 PEMT. 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 PEMT 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.