间接免疫荧光法在小鼠卵子sPLA2 GIID蛋白表达检测中的应用  

Establishment of indirect immunofluorescenct analysis as an assay method for CFTR protein expression in mouse oocytes

作　　者：李红阁[1] 吕建新[1] 陈文颖[2] 石其贤[2] 倪崖[2] 

机构地区：[1]温州医学院,325035 [2]浙江省医学科学院,杭州市310013

出　　处：《实用医学杂志》2011年第7期1141-1144,共4页The Journal of Practical Medicine

基　　金：国家重点基础研究发展规划项目(973计划)(编号:2006CB504000);国家自然科学基金项目(编号:30670788)

摘　　要：目的:建立间接免疫荧光检测小鼠卵子上未知蛋白表达的方法,为实验室开展卵子上未知蛋白表达的研究提供参考。方法:参考间接免疫荧光检测未知蛋白的常用方法,优化实验步骤中各个环节,通过系统性的回顾和分析本实验室近一年来几个阶段对小鼠卵子sPLA2GIID型蛋白表达的探索,建立间接免疫荧光检测小鼠卵子上未知蛋白表达的新方法;并用免疫印迹法(Westernblott)验证结果的准确性。结果:证明小鼠卵子上有很丰富的sPLA2GIID型蛋白的表达;免疫印迹法证实小鼠卵子存在特异的分子质量为16000蛋白条带。结论:本方法测定结果准确可靠、操作简便、重复性好,适宜于实验室推广。Objective To establish indirect immunofluorescence analytic technique to assay mouse oocyte unknown protein expression, and provide a useful reference for laboratory research on oocyte unknown protein expression. Methods Reference to frequent indirect immunofluorescence method to detect unknown protein, optimize steps in the various links, and through systematic survey and analysis of our laboratory work over the past year several stages on mouse oocyte sPLA2 GIID protein expression, thus establish indirect immunofluoreseent new way to assay mouse oocyte unknown protein expression. Results We have proved that a strong sPLA2 GIID protein expression is present on mouse oocyte plasma membrane and detectes a band of 16 kD protein. Conclusions The results demonstrated that the method is accurate, reliable, easy to operate, repetitive and suitable for laboratory purposes.

关 键 词：小鼠卵子 超促排卵 sPLA2GIID蛋白 间接免疫荧光 

分 类 号：R-331[医药卫生]