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肌酸激酶分子是由脑型亚单位(B)和肌型亚单位(M)组成的二聚体。正常组织中常含3种同工酶,按电泳快慢顺序分别为CK-BB(CK1)、CK-MB(CK2)和CK-MM(CK3)。CK-MB主要存在于心肌组织中,占血清中CK的0~4%;CK-MB有自己的亚型,包括MB2(组织形式)和MB1(转化形式)。CK和CK-MB是心肌组织损伤的敏感指标之一,特别是CK-MB更具特异性。心肌损伤过程中CK的升高主要是由于CK-MB的升高引起,表现为二者的同时升高。 本试剂盒采用双抗体夹心法酶联免疫吸附试验(ELISA)。往预先包被有小鼠肌酸激酶MB同工酶(CKMB)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肌酸激酶MB同工酶(CKMB)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 CKMB. 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 CKMB 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.