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巨噬细胞集落刺激因子(M-CSF)是一种分泌型细胞因子,是一种参与细胞信号传导的较小的蛋白质。可使造血干细胞分化为巨噬细胞或其他相关细胞类型。真核细胞也产生M-CSF,以对抗细胞间的病毒感染。它是实验中描述的三种集落刺激因子之一。M-CSF与集落刺激因子1受体结合。该蛋白的活性形式以二硫键连接的同源二聚体形式存在于细胞外,并被认为是由膜结合的前体的蛋白水解产生的。已发现该基因有四个转录变体,编码三种不同的异构体(一种蛋白多糖、糖蛋白和细胞表面蛋白)。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有人巨噬细胞集落刺激因子(M-CSF)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巨噬细胞集落刺激因子(M-CSF)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 M-CSF. 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 M-CSF 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.