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细胞毒性T淋巴细胞相关抗原4(CTLA-4),也被称为CD152(分化簇152),是一种蛋白受体,作为免疫检查点发挥作用,并下调免疫反应。CTLA-4在调节性T细胞中是组成性表达的,但只有在激活后才在常规T细胞中上调。CTLA-4蛋白由小鼠的Ctla-4基因编码。它是免疫球蛋白超家族的一个成员,由活化的T细胞表达,并向T细胞传递抑制性信号。CTLA-4与T细胞共刺激蛋白CD28同源,两种分子都与抗原呈递细胞上的CD80和CD86结合。它向T细胞传递抑制性信号,而CD28传递的是刺激性信号。通过T细胞受体和CD28激活T细胞会导致CTLA-4的表达增加。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠细胞毒性T淋巴细胞相关抗原4(CTLA-4;CD152)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细胞毒性T淋巴细胞相关抗原4(CTLA-4;CD152)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 CTLA-4;CD152. 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 CTLA-4;CD152 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.