Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Proton pump inhibitors are associated with reduced periodontal disease severity.

JADA foundational science·2026
Same author

Publisher Correction: Dynamic realization of emergent high-dimensional optical vortices.

Nature communications·2026
Same author

Dynamic realization of emergent high-dimensional optical vortices.

Nature communications·2025
Same author

Generation of site-specific ubiquitinated histones through chemical ligation to probe the specificities of histone deubiquitinases.

Frontiers in epigenetics and epigenomics·2025
Same author

Trafficking of K63-polyubiquitin-modified membrane proteins in a macroautophagy-independent pathway is linked to ATG9A.

Molecular biology of the cell·2025
Same author

Efficacy of Olmesartan/Amlodipine Single-Pill Combination on 24-h Mean Systolic Blood Pressure Measured by Ambulatory Monitoring in Non-Responders to Valsartan or Candesartan Monotherapy.

Journal of clinical hypertension (Greenwich, Conn.)·2024
Same journal

Mapping the 3D Chromosome Organization of a Biosynthetic Gene Cluster by Capture Hi-C (CHi-C).

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Mapping the 3D Chromosome Organization of Streptomyces by Hi-C.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

CUT&Tag Epigenomic Profiling of Biosynthetic Gene Clusters in Arabidopsis thaliana.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Rhizobium rhizogenes-Mediated Hairy Root Transformation Protocol for Lotus japonicus and Other Legumes.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Characterization of Bioactive Saponins from Sea Cucumbers.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for Functional Validation of Terpenoid Metabolic Clusters in Nicotiana benthamiana and Aspergillus oryzae.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Aug 22, 2025

Measuring Enzymatic Activity of Neurodevelopmental Disorder-Associated Deubiquitylating Enzymes via an In Vitro Ubiquitin Chain Cleavage Assay
07:05

Measuring Enzymatic Activity of Neurodevelopmental Disorder-Associated Deubiquitylating Enzymes via an In Vitro Ubiquitin Chain Cleavage Assay

Published on: September 27, 2024

528

Real-Time Deubiquitination Assays Using a Free Ubiquitin Sensor.

Yun-Seok Choi1, Robert E Cohen2

  • 1School of Natural Sciences, Black Hills State University, Spearfish, SD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed a new sensor, Atto532-tUI, to monitor deubiquitinating enzyme (DUB) activity in real time. This tool enables the study of DUBs using various ubiquitin conjugates, advancing deubiquitination assays.

Keywords:
DUB assayDeubiquitinaseDeubiquitinationFluorescent ubiquitin sensorUbiquitintUI

More Related Videos

Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
09:45

Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples

Published on: May 10, 2015

9.8K
Detection of Protein Ubiquitination
09:00

Detection of Protein Ubiquitination

Published on: August 19, 2009

43.0K

Related Experiment Videos

Last Updated: Aug 22, 2025

Measuring Enzymatic Activity of Neurodevelopmental Disorder-Associated Deubiquitylating Enzymes via an In Vitro Ubiquitin Chain Cleavage Assay
07:05

Measuring Enzymatic Activity of Neurodevelopmental Disorder-Associated Deubiquitylating Enzymes via an In Vitro Ubiquitin Chain Cleavage Assay

Published on: September 27, 2024

528
Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
09:45

Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples

Published on: May 10, 2015

9.8K
Detection of Protein Ubiquitination
09:00

Detection of Protein Ubiquitination

Published on: August 19, 2009

43.0K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Deubiquitinating enzymes (DUBs) are crucial for cellular processes, removing ubiquitin (Ub) from various substrates.
  • Current DUB activity assays often rely on artificial substrates, limiting their applicability.
  • Real-time monitoring of DUBs with native substrates is essential for accurate activity assessment.

Purpose of the Study:

  • To develop a novel sensor for real-time detection of free ubiquitin release.
  • To establish a versatile method for monitoring deubiquitinating enzyme activity using diverse ubiquitin conjugates.
  • To present protocols for the preparation and application of the Atto532-tUI sensor.

Main Methods:

  • Preparation of Atto532-tUI, a high-affinity sensor for free ubiquitin.
  • Utilizing the sensor in real-time deubiquitination assays.
  • Monitoring the release of free ubiquitin from various ubiquitin conjugates.

Main Results:

  • The Atto532-tUI sensor effectively detects free ubiquitin release in real time.
  • The developed assay allows monitoring of deubiquitinating enzyme activity with various ubiquitin conjugates.
  • Protocols for sensor preparation and assay implementation are provided.

Conclusions:

  • Atto532-tUI is a valuable tool for real-time deubiquitination assays.
  • This method offers a versatile alternative to traditional DUB activity assays.
  • The developed protocols facilitate the study of DUBs and their substrates.