摘要
目的快速检测血液中脂联素(Acrp30)的含量。方法大肠杆菌表达Acrp30蛋白的球状区域gAcrp30,用gAcrp30重组蛋白免疫新西兰家兔,获得滴度为10 000的一抗。采用竞争酶联免疫吸收方法(ELISA),将抗原包被在酶标板上,一抗和经过一系列稀释的标准抗原-抑制物或者是待测样品预先结合30 min,再和酶标板上的抗原反应,继而加入二抗(羊抗兔),显色,读取OD450值。以标准抗原的稀释倍数的对数为横坐标,吸光度为纵坐标,绘制标准抑制曲线。结果待测样品的浓度即可从标准曲线上查出。结论建立了一种快速检测体内脂联素含量的方法。
Objective Recent research indicated that Acrp30 is related with obesity, diabetes and atherosclerotic. Quickly detecting the content of plasma Acrp30 is of important significance clinically. Methods Here, full- length Acrp30 and its C- terminal globular head domain （gAcrp30） were expressed in Escherichia coli and gAcrp30 was used to immunize a rabbit to obtain polyclonal antiserum with titer of 10 000. Competitive Enzyme - Linked Immunosorbent Assay （ELISA） was used in the experiment, the standard antigen was covered in each well, secondly, the antibody of gAcrp30 was mixed with standard antigen（ as a stayer） or samples for 30 minutes, then the mixture was added to each well, after that the second antibody was added, finally TMD added, after the reaction was stopped by 2 mol/L H2 SO4, read at 450 mn immediately. A standard graph was plotted by using the logarithm of standard antigen diluted times as X - axis, and the OD450 as Y - axis. Results The content of plasma sample can be detected with the graph. Conclusion A method of quickly detecting the content of plasma Acrp30 was founded.
出处
《南华大学学报（医学版）》
CAS
2006年第1期37-39,43,共4页
Journal of Nanhua University(Medical Edition)
