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NF-κB(或NF-kappaB,"激活B细胞的核因子卡帕光链增强剂")是一种控制DNA转录、细胞因子产生和细胞生存的蛋白质复合物。它几乎存在于所有动物细胞类型中,并参与细胞对刺激的反应,如压力、细胞因子、自由基、重金属、紫外线照射、氧化的低密度脂蛋白以及细菌或病毒抗原。NF-κB在调节对感染的免疫反应中起着关键作用,它还与突触可塑性和记忆的过程有关,与B细胞中免疫球蛋白轻链增强子的11个碱基对序列相互作用。NF-κB家族的所有蛋白在其N端都有一个Rel同源结构域。NF-κB蛋白的一个亚家族,包括RelA、RelB和c-Rel,在其C端有一个转录域。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有大鼠核因子κB(NF-κB)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核因子κB(NF-κB)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 NF-κB. 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 NF-κB 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.