Two new monoclonal antibodies for biochemical and flow cytometric analyses of human interferon regulatory factor-3 activation, turnover, and depletion

Methods. 2013 Feb;59(2):225-32. doi: 10.1016/j.ymeth.2012.05.011. Epub 2012 Jun 13.

Abstract

Interferon regulatory factor-3 (IRF-3) is a master transcription factor that drives the host intracellular innate immune response to virus infection. The importance of IRF-3 in innate immune responses is highlighted by the fact that pathogenic viruses have developed strategies for antagonism of IRF-3. Several tools exist for evaluation of viral regulation of IRF-3 activation and function, but high-quality monoclonal antibodies that mark the differential activation states of human IRF-3 are lacking. To study IRF-3 activation, turnover, and depletion in a high-throughput manner in the context of virus infection, we have developed two new monoclonal antibodies to human IRF-3. These antibodies detect IRF-3 in virus-infected cells in a wide variety of assays and provide a new tool to study virus-host interactions and innate immune signaling.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal, Humanized / genetics*
  • Antibodies, Monoclonal, Humanized / metabolism*
  • Cell Line, Tumor
  • Cells, Cultured
  • Flow Cytometry / methods*
  • HeLa Cells
  • Host-Pathogen Interactions / immunology
  • Humans
  • Interferon Regulatory Factor-3 / antagonists & inhibitors
  • Interferon Regulatory Factor-3 / immunology*
  • Interferon Regulatory Factor-3 / metabolism*
  • Mice
  • Molecular Sequence Data

Substances

  • Antibodies, Monoclonal, Humanized
  • IRF3 protein, human
  • Interferon Regulatory Factor-3