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Updated: May 15, 2026

Time-resolved Förster Resonance Energy Transfer Assays for Measurement of Endogenous Phosphorylated STAT Proteins in Human Cells
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Flow cytometric analysis of STAT phosphorylation.

Jane Murphy1, Gabrielle L Goldberg

  • 1Reid Rheumatology Laboratory, Inflammation Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|January 9, 2013
PubMed
Summary

Flow cytometry enables single-cell analysis of STAT phosphorylation. This method uses specific antibodies to detect phosphorylated STAT proteins, allowing detailed study in diverse cell populations or within single cell types.

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Area of Science:

  • Cellular signaling
  • Immunology
  • Molecular biology

Background:

  • Signal transducer and activator of transcription (STAT) proteins are key mediators of cellular signaling.
  • STAT phosphorylation is a critical step in their activation and downstream effects.
  • Understanding STAT phosphorylation dynamics is essential for deciphering cellular responses.

Purpose of the Study:

  • To demonstrate the utility of flow cytometry for analyzing STAT phosphorylation.
  • To enable the study of STAT phosphorylation at a single-cell level.
  • To investigate STAT phosphorylation within heterogeneous cell populations and single cell types.

Main Methods:

  • Cells are fixed and permeabilized to allow antibody access.
  • Staining with specific antibodies targeting phosphorylated STAT proteins.
  • Analysis using flow cytometry to quantify STAT phosphorylation on a per-cell basis.

Main Results:

  • Flow cytometry successfully quantifies STAT phosphorylation in individual cells.
  • The method allows for the study of STAT phosphorylation in heterogeneous cell mixtures.
  • Single STAT phosphorylation can be analyzed within a single cell type.

Conclusions:

  • Flow cytometry is a powerful tool for studying STAT phosphorylation.
  • This technique provides high-resolution insights into STAT signaling pathways.
  • The method facilitates the investigation of STAT activation in complex biological systems.