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

RFFL-mediated protein quality control limits functional rescue of TRID-CFTR modulator combination therapy for cystic fibrosis nonsense mutations.

Cellular and molecular life sciences : CMLS·2026
Same author

Antisense oligonucleotide targeting the E3 ligase RFFL potentiates CFTR modulator efficacy in CF primary bronchial epithelial cells.

Molecular therapy. Nucleic acids·2025
Same author

Ca<sup>2+</sup>-driven PDIA6 biomolecular condensation ensures proinsulin folding.

Nature cell biology·2025
Same author

RFFL inhibition increases cell surface CFTR and reduces IL-8 production in airway epithelial cells upon COPD-associated environmental pathogen exposure.

Biochemical and biophysical research communications·2025
Same author

Perturbation of EPHA2 and EFNA1 <i>trans</i> binding amplifies inflammatory response in airway epithelial cells.

iScience·2025
Same author

Interaction of a pyrene derivative with cationic [60]fullerene in phospholipid membranes and its effects on photodynamic actions.

Beilstein journal of organic chemistry·2024

Related Experiment Video

Updated: Nov 15, 2025

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates
09:47

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates

Published on: May 10, 2022

2.9K

ELISA Based Protein Ubiquitylation Measurement.

Yuka Kamada1, Ryosuke Fukuda1, Tsukasa Okiyoneda1

  • 1Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.

Bio-Protocol
|March 3, 2021
PubMed
Summary

This study presents a simple ELISA protocol to quantify protein ubiquitylation. This method allows for easy measurement of ubiquitylation for biotin-tagged proteins, improving upon traditional techniques.

Keywords:
CFTRELISAMembrane proteinPoly-ubiquitylationProtein degradationUbiquitin

More Related Videos

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
11:54

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Published on: March 23, 2020

10.0K
Detection of Protein Ubiquitination
09:00

Detection of Protein Ubiquitination

Published on: August 19, 2009

43.3K

Related Experiment Videos

Last Updated: Nov 15, 2025

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates
09:47

Evaluation of Substrate Ubiquitylation by E3 Ubiquitin-ligase in Mammalian Cell Lysates

Published on: May 10, 2022

2.9K
Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry
11:54

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Published on: March 23, 2020

10.0K
Detection of Protein Ubiquitination
09:00

Detection of Protein Ubiquitination

Published on: August 19, 2009

43.3K

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cellular Biology

Background:

  • Ubiquitylation is a critical post-translational modification regulating protein degradation via proteasomal and lysosomal pathways.
  • Current methods for measuring ubiquitylation, like immunoblotting, often require complex protein isolation techniques such as denaturing immunoprecipitation.
  • Accurate quantification of ubiquitylation is essential for understanding cellular processes and disease mechanisms.

Purpose of the Study:

  • To develop and present a straightforward protocol for quantifying protein ubiquitylation.
  • To offer an alternative, accessible method for researchers to measure ubiquitylation levels.
  • To facilitate the study of ubiquitylation in various biological contexts.

Main Methods:

  • The protocol utilizes an Enzyme-Linked Immunosorbent Assay (ELISA) format.
  • Proteins of interest are tagged with biotin to enable specific detection.
  • Quantification is achieved by measuring the ubiquitylation of the biotin-tagged protein-of-interest.

Main Results:

  • The developed ELISA protocol provides an easy and efficient way to quantify protein ubiquitylation.
  • This method simplifies the measurement process compared to traditional immunoprecipitation and immunoblotting techniques.
  • The protocol is suitable for biotin-tagged proteins, allowing for targeted ubiquitylation analysis.

Conclusions:

  • The described ELISA protocol offers a user-friendly approach to quantifying protein ubiquitylation.
  • This method can be widely adopted by researchers for routine analysis of ubiquitylation.
  • The protocol enhances the accessibility of ubiquitylation studies in molecular and cellular biology.