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髓鞘碱性蛋白(MBP)是一种被认为在神经系统中的神经髓鞘化过程中很重要的蛋白质。髓鞘是神经系统特有的多层膜,其功能是作为绝缘体,大大增加轴突冲动传导的速度。MBP维持髓鞘的正确结构,与髓鞘膜中的脂类相互作用。该蛋白定位在中枢神经系统和造血系统的各种细胞上。中枢神经系统中的MBP库非常多样化,有几种剪接变体被表达,包括磷酸化、甲基化、脱酰胺化和瓜氨酸化。这些形式因序列中各种内部位置的短(10到20个残基)肽的存在或不存在而不同。而MBP的主要形式是一个大约18.5Kd(170个残基)的蛋白质。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有大鼠髓鞘碱性蛋白(MBP)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大鼠髓鞘碱性蛋白(MBP)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 Rat MBP. 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 Rat MBP 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.