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

Tobemstomig, a Novel Bispecific Antibody, Preferentially Blocks PD-1 and LAG-3 on CD8 TILs to Expand Stem-like T Cells for Sustained Tumor Control.

Cancer research communications·2026
Same author

A kinetics-based model of haematopoiesis reveals extrinsic regulation of skewed lineage output from stem cells.

Nature cell biology·2026
Same author

Recurrent DNA break clusters drive replication-stress-induced copy number variants and genome diversification.

Nature communications·2026
Same author

Nuclear export block of tyrosine-phosphorylated STAT1 facilitates rapid responsiveness to interferon-γ stimuli at the expense of signal strength.

Biochimica et biophysica acta. Gene regulatory mechanisms·2026
Same author

Metabolite-induced DNA damage drives stochastic stem cell loss and clonal hematopoiesis.

Cell stem cell·2026
Same author

An mTORC2-Lipid Signaling Axis Controls Stress-Induced Organismal Death.

Research square·2026

Related Experiment Video

Updated: Mar 2, 2026

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
09:07

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay

Published on: December 19, 2018

6.9K

A sensitive, semi-quantitative mammalian two-hybrid assay.

Elisabeth Riegel1, Thomas Heimbucher2, Thomas Höfer3

  • 1Department of Applied Life Sciences, University of Applied Sciences, FH Campus Wien, Vienna, Austria.

Biotechniques
|May 23, 2017
PubMed
Summary

We developed an inducible mammalian two-hybrid (iM2H) system to improve the study of protein-protein interactions. This enhanced system offers significantly higher sensitivity and allows for semi-quantitative assessment of interaction strengths.

Keywords:
mammalian two-hybridprotein–protein interactions

More Related Videos

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

14.3K
Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen
11:30

Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen

Published on: January 20, 2017

12.1K

Related Experiment Videos

Last Updated: Mar 2, 2026

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay
09:07

Detecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay

Published on: December 19, 2018

6.9K
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

14.3K
Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen
11:30

Unravelling the Function of a Bacterial Effector from a Non-cultivable Plant Pathogen Using a Yeast Two-hybrid Screen

Published on: January 20, 2017

12.1K

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Protein-protein interactions are essential for cellular functions.
  • Mammalian two-hybrid (M2H) systems are used to study these interactions in a physiological context.
  • Conventional M2H systems have limitations in sensitivity and quantifying interaction strength.

Purpose of the Study:

  • To develop a more sensitive and quantitative mammalian two-hybrid system.
  • To create an inducible system for better control over protein interactions.
  • To enable semi-quantitative assessment of protein-protein interaction strengths.

Main Methods:

  • Development of an inducible mammalian two-hybrid (iM2H) system.
  • Utilized a small-molecule dimerizer (rapalog) for recruitment of transactivation domains.
  • Incorporated internal reference interactions for semi-quantitative analysis.
  • Tested the system with Groucho/Tle proteins and their binding partners.
  • Evaluated drug screening applicability by testing a small-molecule inhibitor.

Main Results:

  • The iM2H system demonstrated up to a 100-fold increase in sensitivity compared to conventional M2H.
  • The system allows for semi-quantitative assessment of protein interaction strengths using reference interactions.
  • Successfully applied the iM2H system to study established protein networks.
  • Demonstrated the utility of iM2H for drug screening by detecting inhibitor effects on protein interactions.

Conclusions:

  • The inducible mammalian two-hybrid (iM2H) system significantly enhances sensitivity and quantitation of protein-protein interactions.
  • iM2H provides an improved platform for studying mammalian protein interactions and their networks.
  • The system is valuable for both fundamental research and drug discovery applications.