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甲型肝炎病毒细胞受体 2(HAVCR2)也被称为 T 细胞免疫球蛋白粘蛋白分 子 3(TIM-3),是一种 HAVCR2(TIM-3)基因编码的蛋白质。它是 T 淋巴细 胞(CD4+和 CD8+T 细胞)、骨髓细胞(单核细胞、巨噬细胞、DC、肥大 细胞、NK 细胞)或不同类型肿瘤的各种细胞的一种跨膜蛋白。该受体是一 个免疫检查点,与其他抑制性受体包括细胞程序性死亡蛋白 1(PD-1)和淋 巴细胞激活基因 3 蛋白(LAG3)一起介导 CD8+T 细胞在增殖和分泌细胞因 子如 TNF-α、IFN-γ和 IL-2 方面的衰竭。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人T细胞免疫球蛋白粘蛋白分子3(TIM-3)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人T细胞免疫球蛋白粘蛋白分子3(TIM-3)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 Human TIM-3. 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 Human TIM-3 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.