Detection of intracellular phosphorylated STAT-1 by flow cytometry

Thomas A. Fleisher, Susan E. Dorman, Jill A. Anderson, Michael Vail, Margaret R. Brown, Steven M. Holland

Research output: Contribution to journalArticlepeer-review

Abstract

We have applied flow cytometry to the investigation of interferon-γ activation of human monocytes. This approach uses monoclonal antibodies that distinguish between the native and phosphorylated forms of STAT-1. It enables rapid and quantitative assessment of STAT-1 phosphorylation on a discrete cell basis and is both more sensitive and less time consuming than immunoblotting. Furthermore, it allows for discrimination between a mixture of cells that differ in their response to interferon-γ. This approach should allow for the evaluation of different intracellular signaling pathways using a combination of monoclonal reagents that are specific for native and activation modified proteins. Application of this form of testing should prove valuable in screening for signaling defects in selected patients with recurrent infections. In addition, this technique should permit dissection of a full range of cellular signaling pathways at the protein level.

Original languageEnglish (US)
Pages (from-to)425-430
Number of pages6
JournalClinical Immunology
Volume90
Issue number3
DOIs
StatePublished - Mar 1999

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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