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BioReagent 级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
肽酰甘氨酸α酰胺化单加氧酶,也被称为PAM或PAL,是由PAM基因编码的酶。该酶参与许多信号肽和某些脂肪酸酰胺的生物合成。这种转化是通过将激素原转化为相应的酰胺来实现的。该酶是产生肽酰胺的唯一已知途径,其使肽更具亲水性。它具有两个具有催化活性的酶活性结构域 - 肽基甘氨酸α-羟基化单加氧酶(PHM)和肽基-α-羟基甘氨酸α-酰胺裂解酶(PAL)。这些催化结构域依次工作,将神经内分泌肽催化成活性α-酰胺产物。它催化的反应途径通过量子隧穿和底物预制进入。已经描述了编码不同亚型的多个交替剪接的转录本变体,但它们的一些全长序列尚不清楚。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人肽酰甘氨酸α酰胺化单加氧酶(PAM)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肽酰甘氨酸α酰胺化单加氧酶(PAM)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 PAM. 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 PAM 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.