贵州医科大学学报

2013, (01) 24-26

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免疫磁珠法分离preS1结合蛋白
Isolation of Hepatitis B Virus PreS1 Binding Protein by Immunomagnetic Beads

杨丽;王良宏;潘卫;李兴;杨国珍;
YANG Li,WANG Lianghong,PAN Wei,LI Xing,YANG Guozheng(Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China)

摘要(Abstract):

目的:用免疫磁珠分离人肝癌细胞(HepG2)细胞膜上与乙型肝炎病毒(HBV)preS1肽段结合的蛋白,建立快速分离膜受体蛋白的方法。方法:提取HepG2细胞膜蛋白,以生物素标记的preS1肽段为钓饵,与提取的膜蛋白4℃孵育过夜,加入链霉亲和素标记的磁珠,洗涤磁珠后收集结合蛋白,通过SDS-PAGE电泳分离结合蛋白。结果:用磁珠的方法所收集到的结合蛋白,通过SDS-PAGE电泳分离出了多条条带,比较明显的45~97 ku有9条,>97 ku处有5条,<20 ku处有2条。结论:通过免疫磁珠分离法可有效的获得HepG2细胞膜上preS1肽段的结合蛋白。
Objective: Using immunomagnetic beads method to separate the receptor-binding protein of HepG2 membrane that binding hepatitis B virus preS1 region,and to establish a fast method for separating membrane receptor proteins.Methods: Membrane protein was extracted from HepG2 cells,then combined with preS1 peptide which labeled with biotin,incubated overnight at 4 ℃ with the extraction of membrane proteins.After that,streptavidin-conjugated beads were added.After washing the beads and collection of binding proteins,proteins were separated by SDS-PAGE electrophoresis.Results: Binding proteins were collected by immunomagnetic beads and multiple stripes were separated by SDS-PAGE electrophoresis.There were nine stripes between 45 ku~97 ku obviously,and five stripes in area lager than 97 ku,two stripes in area smaller than 20 ku.Conclusions: The preS1 peptide binding protein of HepG2 membrane can be effectively obtained by using immunomagnetic beads separation method.

关键词(KeyWords): 肝炎病毒,乙型;免疫磁化分离;生物素;亲和素
hepatitis B virus;immunomagnetic separation;biotin;avidin

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基金项目(Foundation): 贵州省科学技术基金,黔科合J字[2008]2157;; 贵州省优秀人才省长专项基金(2005)297

作者(Author): 杨丽;王良宏;潘卫;李兴;杨国珍;
YANG Li,WANG Lianghong,PAN Wei,LI Xing,YANG Guozheng(Department of Clinical Laboratory Science,Guiyang Medical College,Guiyang 550004,Guizhou,China)

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