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DNA(胞嘧啶-5)甲基转移酶 1 是一种催化甲基转移到 DNA 中特定 CpG 结构的酶,这一过程称为 DNA 甲基化。它由 DNMT1 基因编码。DNMT1 是 DNA 甲基转移酶家族的一部分,该家族主要由 DNMT1、DNMT3A 和DNMT3B 组成。这种酶负责维持 DNA 甲基化,从而确保这种表观遗传模式在细胞分裂中的保真度。然而,Dnmt1 可以在特定的基因组环境中催化新的DNA 甲基化,包括可转座元素和父本印记控制区。与 DNMT1 相关的疾病包括小脑共济失调、耳聋和嗜睡症,常染色体显性和神经病变,遗传性感觉, Ie 型。 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DNA甲基转移酶1(DNMT1)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DNA甲基转移酶1(DNMT1)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 DNMT1. 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 DNMT1 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.