| 胡小波,颜志强,陈霞,杨胜利,许金华,龚毅.竞争ELISA法快速检测血液中脂联素的含量.[J].中南医学科学杂志.,2006,(1):37-39,43. |
| 竞争ELISA法快速检测血液中脂联素的含量 |
| Quickly Detecting the Content of Plasma Acrp30 by Competitive ELISA |
| 投稿时间：2005-10-22 |
| DOI： |
| 中文关键词: 脂联素 竞争ELISA 肥胖 糖尿病 动脉硬化 |
| 英文关键词:Adiponectin competitive ELISA obesity diabetes atherosclerotic |
| 基金项目: |
| 胡小波 颜志强 陈霞 杨胜利 许金华 龚毅 |
| [1]中科院上海生命科学研究院,上海200233 [2]南华大学生化教研室,上海200233 |
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| 中文摘要: |
| 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. |
| 英文摘要: |
| 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. |
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