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中性粒细胞胞外诱捕网络(NETs)是细胞外纤维的网络,主要由中性粒细胞的DNA 组成,它与病原体结合。中性粒细胞是免疫系统对抗感染的第一道防线, NETs 使中性粒细胞能够杀死细胞外病原体,同时最大限度地减少对宿主细胞的损害。在体外用药理学制剂乙酸苯丙醇酯(PMA)、白细胞介素 8(IL-8) 或脂多糖(LPS)激活后,中性粒细胞释放颗粒蛋白和染色质,通过一个主动过程形成称为 NET 的细胞外纤维基质。NETs 在炎症部位大量产生,并在局部提供高浓度的抗菌分子,捕获并杀死各种病原体,快速控制病菌在体内感染,起到免疫抗菌。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人中性粒细胞胞外诱捕网络(NETs)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中性粒细胞胞外诱捕网络(NETs)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 NETs. 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 NETs 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.