中华临床免疫和变态反应杂志2017，Vol.11Issue(1)：1-5,5.DOI:10.3969/j.issn.1673-8705.2017.01.001
双向电泳结合免疫印迹技术分析蜉蝣变应原
Analysis of Allergenic Components of Ephemeroptera with Two DimensionalElectrophoresis Followed by Immunoblot
摘要
Abstract
Objective To investigate the major allergens of Ephemeroptera based on the analysis of allergenic components of its protein extract by using two dimensional electrophoresis (2-DE) and immunoblot(Western blot).Methods Ephemeroptera protein extract was prepared by routine method.Allergenic components of Ephemeroptera were analyzed by using 2-DE and immunoblot.The Ephemeroptera allergic patients` serum was used as first antibody, and mouse anti-human IgE antibody as second antibody.Results About 805 soluble proteins spots were identified by using two dimensional electrophoresis with Coomassie brilliant blue staining.The calculated molecular weight of the most identified proteins ranged from 20 000-130 000, and the calculated pI ranged from 4.0-9.5.Twenty-seven allergenic spots were identified using 2-DE and immunoblot compared with the control group.Conclusion 2-DE is a good method for analysis of Ephemeroptera protein components and further investigation on its allergenic components.关键词
蜉蝣/变应原/双向电泳/免疫印迹Key words
Ephemeroptera/allergen/two dimensional electrophoresis/Western blot分类
医药卫生引用本文复制引用
刘硕,蔡笃程,李春林,孟光,龙绮,谢伟伟..双向电泳结合免疫印迹技术分析蜉蝣变应原[J].中华临床免疫和变态反应杂志,2017,11(1):1-5,5.基金项目
国家自然科学基金(81160129),海南省自然科学基金(30871,811145),海南省国际科技合作重点计划项目(2012-GH010),海口市重点科技计划项目(海科立2007-0080号,海科工信立[2012]72号),海南省卫生厅科研项目(琼卫2007-79) （81160129）
