竞争ELISA法快速检测血液中脂联素的含量

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.

Quickly Detecting the Content of Plasma Acrp30 by Competitive ELISA

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 H_2SO_4,read at 450 nm immediately.A standard graph was plotted by using the logarithm of standard antigen diluted times as X-axis,and the OD_(450) 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.