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β-内啡肽(β-EP)是一种内源性阿片类神经肽和肽类激素,在中枢神经系统和外周神经系统的某些神经元中产生。内啡肽存在于下丘脑的神经元,以及垂体的神经元。它来源于β-促肾上腺皮质激素,后者在垂体中由较大的多肽前体--原肾上腺皮质激素(POMC)产生。POMC被裂解为两种神经肽,促肾上腺皮质激素(ACTH)和β-促肾上腺皮质激素。然后,β-内啡肽的形成是β-促脂素C端区域裂解的结果,产生一个31个氨基酸长的神经肽,具有α-螺旋二级结构。然而,POMC也产生其他肽类激素,包括α-和γ-黑素细胞刺激素(MSH),是由被称为原激素转化酶的内部酶在细胞内加工产生的。 本试剂盒采用竞争法酶联免疫吸附试验(ELISA)。往预先包被有人β-内啡肽(β-EP)抗原的微孔中,依次加入样本、标准品、Biotin标记的抗体,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β-内啡肽(β-EP)呈负相关。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 competitive enzyme-linked immunosorbent assay (ELISA) principle. Microplate wells are pre-coated with Human β-EP antigen. The sample, standards, a biotin-labeled antibody, and an HRP enzyme conjugate are then added sequentially to the wells. The procedure includes incubation and wash steps between each addition. TMB substrate is used for color development. In the presence of the peroxidase (HRP) enzyme, TMB is catalyzed to produce a blue color, which is then converted into a final stable yellow color upon the addition of an acid stop solution. The intensity of the resulting yellow color is inversely proportional to the concentration of Human β-EP in the sample. The optical density (OD) is measured at a wavelength of 450 nm using a microplate reader, and the concentration of the target analyte in the sample is calculated based on the standard curve.