用多克隆抗体检测干扰素调节因子7的表达(英文)  

Generation of Polyclonal Antibody against Interferon Regulatory Factor 7

作　　者：刘晓辉[1] 秦宇[2] 马星[1] 齐婧[1] 王金忠[1] 耿运琪[1] 

机构地区：[1]南开大学泰达生物技术学院分子微生物学与技术教育部重点实验室,天津300071 [2]天津医科大学基础医学院,天津300070

出　　处：《南开大学学报（自然科学版）》2010年第1期44-47,共4页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：Supported by National Natural Science Foundation of China(30870129);Doctoral Fund of Ministry of Education(200800550021)

摘　　要：干扰素调节因子7(IRF-7)是调节Ⅰ型干扰素依赖型先天免疫反应的关键因子,在真核细胞防御反应中发挥重要作用。本文在大肠杆菌中表达了重组IRF-7,利用亲和层析的方法进行了纯化,并制备了鼠抗IRF-7的多克隆抗体。所得到的抗血清能够在稀释27 000倍后成功用于免疫印迹检测。该抗血清不但能识别来源于大肠杆菌的抗原,还能检测真核细胞内转染后表达的IRF-7。使用该抗体证实,转染293T细胞后表达的IRF-7主要分布在细胞质内。所制备的抗血清可用于IRF-7的表达和功能研究。Interferon regulatory factor 7（IRF-7） is a key regulator of type-Ⅰinterferon-dependent innate immune responses,and plays a critical role in the activation of cellular defense genes.In the present study,recombinant human IRF-7 was expressed in Escherichia coli,purified by affinity chromatography,and utilized to raise polyclonal antibody in BALB/c mice.High-titer antisera were obtained,which successfully detected the antigen at a dilution of 1：27 000 by Western blot analysis.The antisera can recognize not only recombinant IRF-7 expressed in E.coli,but also transfected IRF-7 in mammalian cells.We also showed that the IRF-7 was localized mainly in the cytoplasm of transfected 293T cells.The anti-IRF-7 antibody generated in this study will be useful for the detection of IRF-7 expression and the investigation of IRF-7 function.

关 键 词：干扰素调节因子7 多克隆抗体 免疫印迹 

分 类 号：Q511[生物学—生物化学]