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

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.