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 Experiment Videos

Homogeneous time-resolved IL-2-IL-2R alpha assay using fluorescence resonance energy transfer

K Stenroos1, P Hurskainen, S Eriksson

  • 1Department of Biology, Abo Akademi University, Turku, Finland.

Cytokine
|August 14, 1998
PubMed
Summary

This study introduces a novel homogeneous assay for detecting receptor-ligand interactions using fluorescence resonance energy transfer. The assay efficiently measures binding for interleukin 2 (IL-2) and its receptor, suitable for high-throughput screening.

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

Limited opportunity for nursing students to perform practical nursing skills during clinical placements: a Scandinavian cross-sectional study.

International journal of nursing studies advances·2026
Same author

Adaptation and psychometric evaluation of the Swedish version of the Assessment of Inter professional Team Collaboration Scale (AITCS-S) for use in occupational health services.

Journal of interprofessional care·2022
Same author

Unwanted mainstream nitritation-denitritation causing massive N<sub>2</sub>O emissions in a continuous activated sludge process.

Water science and technology : a journal of the International Association on Water Pollution Research·2021
Same author

Body composition assessed by dual-energy X-ray absorptiometry predicts early infectious complications after liver transplantation.

Journal of human nutrition and dietetics : the official journal of the British Dietetic Association·2016
Same author

Effect of C-H bonds on the quenching of luminescent lanthanide chelates.

Journal of fluorescence·2013
Same author

The role of unfinished root canal treatment in odontogenic maxillofacial infections requiring hospital care.

Clinical oral investigations·2012

Area of Science:

  • Biochemistry
  • Immunology
  • Assay Development

Background:

  • Interleukin 2 (IL-2) plays a crucial role in immune responses.
  • Accurate measurement of IL-2 receptor (IL-2R) interactions is vital for understanding T-cell activation and developing targeted therapies.
  • Existing assays may lack the sensitivity or throughput required for comprehensive analysis.

Purpose of the Study:

  • To develop a novel homogeneous receptor-ligand assay for quantifying IL-2 and IL-2R alpha chain interactions.
  • To utilize fluorescence resonance energy transfer (FRET) for sensitive detection of binding events.
  • To establish an assay suitable for high-throughput screening (HTS) applications.

Main Methods:

  • A homogeneous FRET-based assay was designed using a europium chelate-labeled IL-2 and a Cy5-labeled monoclonal antibody against the IL-2 receptor alpha chain.

Related Experiment Videos

  • Recombinant IL-2 receptor alpha chain membranes were prepared from insect cells infected with baculovirus.
  • Time-resolved fluorometry was employed to measure energy transfer upon excitation of the europium donor.

    • Main Results:

    • The FRET assay demonstrated efficient energy transfer upon successful receptor-ligand complex formation.
    • The assay successfully quantified binding interactions using specific non-neutralizing antibodies.
    • The assay design proved effective for receptor titrations, saturation binding, and competitive binding experiments.

    Conclusions:

    • A rapid, simple, and homogeneous FRET-based assay for IL-2/IL-2R alpha chain interactions has been successfully developed.
    • This assay is particularly well-suited for high-throughput screening of receptor-ligand binding.
    • The methodology offers a valuable tool for immunological research and drug discovery.