Generation of Polyclonal Antibody against Interferon Regulatory Factor 7

Interferon regulatory factor 7 (IRF-7) is a key regulator of type-I interferon-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.