title: [Preparation and identification of the polyclonal antibody against extracellular domain (405T-573Q) of rat Toll-like receptor 2].
abstract: OBJECTIVE
  To prepare the polyclonal antibody of the extracellular domain (405T-573Q) of rat Toll-like receptor 2 (TLR2) and identify its activity.
  
  
  METHODS
  Firstly, the extracellular domain (507 bp) of rat TLR2 was amplified from rat hepatocytes by reverse transcription PCR, and the recombinant plasmid pET28a-TLR2ex was constructed. Secondly, the fusion protein TLR2ex-6×His was expressed in E.coli BL21(DE3) and purified by affinity chromatography. And then, the polyclonal antibody was prepared by immunizing the BALB/c mice with the purified fusion protein. Finally, the titer and specificity of the polyclonal antibody were analyzed by ELISA and Western blotting, respectively.
  
  
  RESULTS
  The recombinant plasmid pET28a-TLR2ex was successfully constructed, and the TLR2ex-6×His fusion protein was successfully expressed and purified. Western blotting demonstrated that the antibody from the TLR2ex-6×His-immunized BALB/c mice specifically and selectively bound with rat TLR2. ELISA also indicated that the titer of polyclonal antibody reached 1:76 800.
  
  
  CONCLUSIONS
  The polyclonal antibody against the extracellular domain (405T-573Q) of rat TLR2 has been successfully prepared.
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Lan
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yi
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Fujun
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Shemin
authors.initials: S
authors.lastname: Lv
authors.affiliation: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University, Xi'an 710061, China.
authors.forename: Dongmin
authors.initials: D
authors.lastname: Li
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D051195:Toll-Like Receptor 2 / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; C495343:Tlr2 protein, rat; D051195:Toll-Like Receptor 2