title: Development and characterization of monoclonal antibodies against human aryl hydrocarbon receptor.
abstract: Aryl hydrocarbon receptor (AhR), a ligand-dependent nuclear receptor, is involved in a diverse spectrum of biological and toxicological effects. Due to the lack of three dimensional (3D) crystal or nuclear magnetic resonance structure, the mechanisms of these complex effects of AhR remain to be unclear. Also, commercial monoclonal antibodies (mAbs) against human AhR protein (hAhR), as alternative immunological tools, are very limited. Thus, in order to provide more tools for further studies on hAhR, we prepared two mAbs (1D6 and 4A6) against hAhR. The two newly generated mAbs specifically bound to amino acids 484-508 (located in transcription activation domain) and amino acids 201-215 (located in Per-ARNT-Sim domain) of hAhR, respectively. These epitopes were new as compared with those of commercial mAbs. The mAbs were also characterized by enzyme-linked immunosorbent assay, western blot, immunoprecipitation and indirect immunofluorescence assay in different cell lines. The results showed that the two mAbs could recognize the linearized AhRs in six different human cell lines and a rat hepatoma cell line, as well as the hAhR with native conformations. We concluded that the newly generated mAbs could be employed in AhR-based bioassays for analysis of environmental contaminants, and held great potential for further revealing the spatial structure of AhR and its biological functions in future studies.
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Tian
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Xinhui
authors.initials: X
authors.lastname: Pei
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China. Electronic address: qhxie@rcees.ac.cn.
authors.forename: Heidi Qunhui
authors.initials: HQ
authors.lastname: Xie
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Sherry Li
authors.initials: SL
authors.lastname: Xu
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Jijing
authors.initials: J
authors.lastname: Tian
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Hu
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Haiming
authors.initials: H
authors.lastname: Xu
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Yangsheng
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
authors.forename: Hualing
authors.initials: H
authors.lastname: Fu
authors.affiliation: China Pharmaceutical University, Nanjing 211198, China. Electronic address: zycao1999@hotmail.com.
authors.forename: Zhengyu
authors.initials: Z
authors.lastname: Cao
authors.affiliation: Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China. Electronic address: binzhao@rcees.ac.cn.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D051792:Basic Helix-Loop-Helix Proteins / Q000737:chemistry / Q000276:immunology*; D045744:Cell Line, Tumor; D000939:Epitopes / Q000276:immunology; D006801:Humans; D051379:Mice; D051381:Rats; D018336:Receptors, Aryl Hydrocarbon / Q000737:chemistry / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D051792:Basic Helix-Loop-Helix Proteins; D000939:Epitopes; D018336:Receptors, Aryl Hydrocarbon; C111235:AHR protein, human
keywords: Aryl hydrocarbon receptor; Immunoprecipitation; Indirect immunofluorescence assay; Monoclonal antibody; Western blot