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Protein Dynamics in Living Cells

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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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Updated: May 15, 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

High-throughput flow cytometry compatible biosensor based on fluorogen activating protein technology.

Yang Wu¹, Phillip H Tapia, Gregory W Fisher

¹UNM Center for Molecular Discovery, University of New Mexico School of Medicine, Albuquerque, New Mexico 87131, USA. yawu@salud.unm.edu

Cytometry. Part a : the Journal of the International Society for Analytical Cytology

|January 11, 2013

Summary

This summary is machine-generated.

This study presents a novel biosensor system for tracking multiple trafficking proteins in live cells. The robust and sensitive platform enables detailed mechanistic studies of functional receptor pairs, advancing cellular protein research.

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Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

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Last Updated: May 15, 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

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer

Published on: October 1, 2016

Area of Science:

Cell biology
Biochemistry
Molecular imaging

Background:

Simultaneous monitoring of multiple proteins in live cells is crucial for understanding cellular functions.
Existing fluorescent labeling methods limit mechanistic studies of functional receptor pairs.

Purpose of the Study:

To develop a versatile and sensitive biosensor system for simultaneous monitoring of multiple trafficking proteins.
To overcome limitations of current fluorescent labeling techniques for studying functional receptor pairs.

Main Methods:

Expansion of a hybrid system combining fluorogen-activating protein (FAP) technology and high-throughput flow cytometry.
Utilized human beta2 adrenergic receptor (β2AR) and murine C-C chemokine receptor type 5 (CCR5) as model systems, fused with specific FAP tags.
Characterized receptor function and FAP-fluorogen binding using flow cytometry and confocal microscopy.

Main Results:

Developed a robust, sensitive, and versatile biosensor system for studying multiple trafficking proteins.
Demonstrated highly specific binding between FAP tags and their cognate fluorogens.
Confirmed that FAP-tagged fusion proteins retain similar functionality to their wild-type counterparts.
Successfully employed the system as a counter-screen assay to identify false positives in drug screening.

Conclusions:

The expanded FAP-based biosensor system enables robust and sensitive monitoring of multiple trafficking proteins in live cells.
This technology facilitates mechanistic studies of functional receptor pairs, overcoming limitations of previous methods.
The system proves valuable for drug discovery, particularly in counter-screening assays for receptor regulators.