Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Author Correction: Targeting the SHOC2-RAS interaction in RAS-mutant cancers.

Nature·2025

Same author

Targeting the SHOC2-RAS interaction in RAS-mutant cancers.

Nature·2025

Same author

Reversible Covalent Reactions of Aldehydes and Salicylaldehydes Using a Lysine-Model Substrate.

Chimia·2025

Same author

Discovery of Ligands for TRIM58, a Novel Tissue-Selective E3 Ligase.

ACS medicinal chemistry letters·2023

Same author

Blocking phospholamban with VHH intrabodies enhances contractility and relaxation in heart failure.

Nature communications·2022

Same author

Engineered Cas9 extracellular vesicles as a novel gene editing tool.

Journal of extracellular vesicles·2022

Search research articles

Related Experiment Video

Updated: Jun 6, 2026

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Published on: July 30, 2020

A fluorescence lifetime-based assay for abelson kinase.

Stephan Pritz¹, Gabriele Meder, Klaus Doering

¹Biosyntan GmbH, Berlin, Germany.

Journal of Biomolecular Screening

|December 15, 2010

Summary

This summary is machine-generated.

We developed a new homogeneous assay to measure tyrosine kinase activity using a fluorescent peptide substrate. This method directly tracks enzyme phosphorylation without antibodies, enabling efficient compound profiling.

More Related Videos

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Published on: May 13, 2020

Related Experiment Videos

Last Updated: Jun 6, 2026

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Real-Time Monitoring of Aurora kinase A Activation using Conformational FRET Biosensors in Live Cells

Published on: July 30, 2020

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Visualizing Protein Kinase A Activity In Head-fixed Behaving Mice Using In Vivo Two-photon Fluorescence Lifetime Imaging Microscopy

Published on: June 7, 2019

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Click-Chemistry Based Fluorometric Assay for Apolipoprotein N-acyltransferase from Enzyme Characterization to High-Throughput Screening

Published on: May 13, 2020

Area of Science:

Biochemistry
Enzymology
Assay Development

Background:

Tyrosine kinases are crucial drug targets, necessitating robust methods for activity measurement.
Existing assays often require secondary reagents like antibodies, complicating the process.

Purpose of the Study:

To introduce a novel, homogeneous in vitro assay for quantifying tyrosine kinase enzymatic activity.
To demonstrate a direct, antibody-free method for monitoring substrate phosphorylation.

Main Methods:

Utilized a peptide substrate with a single tyrosine and a cysteine-linked fluorescent probe.
Exploited fluorescence lifetime changes due to dynamic quenching by the nonphosphorylated tyrosine.
Applied the assay to Abelson kinase (c-Abl) for compound profiling.

Main Results:

Successfully demonstrated direct monitoring of tyrosine kinase phosphorylation.
Achieved quantitative determination of enzymatic activity.
Validated the assay for Abelson kinase (c-Abl) and showed potential for other tyrosine kinases.

Conclusions:

The developed assay offers a sensitive and direct approach for tyrosine kinase activity measurement.
This homogeneous assay format simplifies workflows and is adaptable to various tyrosine kinases.
The method is suitable for high-throughput screening and compound profiling in drug discovery.