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Protein Dynamics in Living Cells

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Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: May 25, 2026

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Published on: October 4, 2018

Phospho-specific flow: fixating on the target.

¹Department of Oncology, Johns Hopkins University, Baltimore, MD, USA. levisma@jhmi.edu

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research

|February 4, 2012

Summary

Confirming targeted therapy effectiveness in oncology is crucial. This study demonstrates phospho-specific flow cytometry for real-time, in vivo target inhibition confirmation.

Area of Science:

Oncology
Molecular Biology
Immunology

Background:

Targeted therapies represent a significant advancement in cancer treatment.
A key challenge is verifying target engagement within a living organism (in vivo).
Current methods may lack the precision or real-time capability to confirm target inhibition.

Purpose of the Study:

To present a novel method for confirming in vivo target inhibition.
To demonstrate the utility of phospho-specific flow cytometry in oncology research.
To provide a real-time assessment of targeted therapy efficacy.

Main Methods:

Application of phospho-specific flow cytometry.
In vivo experimental models.
Analysis of protein phosphorylation states as a marker of target activity.

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An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

Related Experiment Videos

Last Updated: May 25, 2026

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Phospho Flow Cytometry with Fluorescent Cell Barcoding for Single Cell Signaling Analysis and Biomarker Discovery

Published on: October 4, 2018

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes

Published on: October 15, 2016

Main Results:

Phospho-specific flow cytometry successfully confirmed target inhibition in vivo.
The method provides real-time data on therapeutic response.
Demonstrated ability to monitor the effects of targeted therapies.

Conclusions:

Phospho-specific flow cytometry is a valuable tool for validating targeted therapies.
This technique enables precise, real-time assessment of in vivo target engagement.
Facilitates rapid decision-making in oncology drug development and clinical practice.