Expression of Recombinant Human FADD,Preparation of Its Polyclonal Antiserum and the Application in Immunoassays  

作　　者：Faiz MMT Marikar 

机构地区：[1]The State Key Laboratory of Pharmaceutical Biotechnology and Jiangsu Center of Hepatobiliary Diseases, Nanjing University

出　　处：《Cellular & Molecular Immunology》2008年第6期471-474,共4页中国免疫学杂志（英文版）

基　　金：financially supported by the following funds to Zi-Chun Hua: the National Nature Science Foundation of China (No. 30425009, 30600320, 30730030, 30330530, 30270291);Jiangsu Provincial Nature Sciences Foundation (No. BK2007715);Jiangsu Provincial Department of Health (No. H200524, H200742)

摘　　要：The wild-type human Fas-associated death domain （FADD） protein was expressed as a His-tag fusion protein in Escherichia coll. Recombinant FADD proteins were purified under the denatured condition. After denatured protein purification, it was refolded and obtained at a yield of about 23 mg/L. Purified FADD exhibited as a homogenous band corresponding to the molecular weight of 31 kDa. Immunization of rabbits against the refolded FADD protein was allowed the production of high titre polyclonal antiserum. This new polyelonal antibody could recognize recombinant FADD protein in Western blot. Immunoreactivity was also observed in immunofiuorescence assay. The low cost polyclonal antiserum was applicable to extensive detection of FADD in various immunoassays. Cellular ＆ Molecular Immunology. 2008;5（6）：471-474.The wild-type human Fas-associated death domain （FADD） protein was expressed as a His-tag fusion protein in Escherichia coll. Recombinant FADD proteins were purified under the denatured condition. After denatured protein purification, it was refolded and obtained at a yield of about 23 mg/L. Purified FADD exhibited as a homogenous band corresponding to the molecular weight of 31 kDa. Immunization of rabbits against the refolded FADD protein was allowed the production of high titre polyclonal antiserum. This new polyelonal antibody could recognize recombinant FADD protein in Western blot. Immunoreactivity was also observed in immunofiuorescence assay. The low cost polyclonal antiserum was applicable to extensive detection of FADD in various immunoassays. Cellular ＆ Molecular Immunology. 2008;5（6）：471-474.

关 键 词：FADD His-tag fusion protein polyclonal antibody immunofluorescence assay 

分 类 号：R392[医药卫生—免疫学]