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题名: Development and characterization of monoclonal antibodies against human aryl hydrocarbon receptor
作者: Tian, Wenjing; Pei, Xinhui; Xie, Heidi Qunhui; Xu, Sherry Li; Tian, Jijing; Hu, Qin; Xu, Haiming; Chen, Yangsheng; Fu, Hualing; Cao, Zhengyu; Zhao, Bin
刊名: JOURNAL OF ENVIRONMENTAL SCIENCES
出版日期: 2016-01
卷号: 39, 期号:0, 页码:165-174
关键词: Aryl hydrocarbon receptor ; Monoclonal antibody ; Western blot ; Immunoprecipitation ; Indirect immunofluorescence assay
DOI: 10.1016/j.jes.2015.11.008
部门归属: 环境化学与生态毒理学国家重点实验室
英文摘要: 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 providemore tools for further studies on hAhR, we prepared twomAbs (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 epitopeswere newas compared with those of commercialmAbs. 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 twomAbs 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. (C) 2015 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V.
收录类别: SCI
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内容类型: 期刊论文
URI标识: http://ir.rcees.ac.cn/handle/311016/35837
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