title: Expression of recombinant human FADD, preparation of its polyclonal antiserum and the application in immunoassays.
abstract: The wild-type human Fas-associated death domain (FADD) protein was expressed as a His-tag fusion protein in Escherichia coli. 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 polyclonal antibody could recognize recombinant FADD protein in Western blot. Immunoreactivity was also observed in immunofluorescence assay. The low cost polyclonal antiserum was applicable to extensive detection of FADD in various immunoassays.
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Center of Hepatobiliary Diseases, Nanjing University, Nanjing, China.
authors.forename: Faiz M M T
authors.initials: FM
authors.lastname: Marikar
authors.forename: Dingyuan
authors.initials: D
authors.lastname: Ma
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Ye
authors.forename: Bo
authors.initials: B
authors.lastname: Tang
authors.forename: Weijuan
authors.initials: W
authors.lastname: Zheng
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Min
authors.initials: M
authors.lastname: Lu
authors.forename: Zichun
authors.initials: Z
authors.lastname: Hua
mesh_terms: D000818:Animals; D002460:Cell Line; D053200:Fas-Associated Death Domain Protein / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D007118:Immunoassay; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology*; D014162:Transfection
chemical_list: D053200:Fas-Associated Death Domain Protein; D007106:Immune Sera; D011994:Recombinant Proteins