摘要
目的建立一种利用二维液相色谱分离和质谱鉴定大鼠附睾液蛋白质组的实验方法,为以后研究其他哺乳动物附睾蛋白质组提供实验基础。方法分离提取大鼠附睾头、体、尾部管腔蛋白,样品经脱盐、浓缩,利用起始缓冲液置换,进行一维色谱聚焦分离,收集pH4．0—8．5之间的组分进行二维反相色谱分离,并通过自动馏分收集器收集分离组份;最后将获得的二维UV图通过ProteoVue软件转换成二维pL／UV图谱;选择含高丰度蛋白的馏分经冷冻干燥浓缩后进行质谱鉴定。结果通过二维液相色谱成功分离了大鼠附睾头、体、尾部附睾液蛋白并建立了二维pL／UV图谱;对二维获得的含高丰度蛋白的馏分进行了质谱鉴定,获得大鼠附睾头、体、尾部主要蛋白。结论为进一步利用二维液相色谱全面分离附睾蛋白和研究附睾功能奠定了基础。
Objective To establish an experimental method of studying rat epididymal fluid proteome by two dimensional liquid chromatography and MALDI-TOF MS and provide a foundation of studying other mammal epididymal proteome. Methods The epididymal fluid proteins extracted were exchanged with start buffer and separated by chromatofocusing in the first dimension. The fractions between pH4. 0 and pH8. 5 collected from 1D were separated by reverse-phase HPLC. The fractions were collected by automatic fraction collector...
出处
《国际遗传学杂志》
CAS
2006年第6期420-423,共4页
International Journal of Genetics
基金
山东省科技发展计划基金(No.032050102)
关键词
蛋白质组
二维液相色谱分离
附睾蛋白质
Proteome
Two dimensional liquid chromatographic fractionation
Epididymal proteins
