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

A fluorescence polarization based Src-SH2 binding assay

B A Lynch1, K A Loiacono, C L Tiong

  • 1ARIAD Pharmaceuticals Inc., Cambridge, Massachusetts 02139, USA.

Analytical Biochemistry
|April 5, 1997
PubMed
Summary
This summary is machine-generated.

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

Architecture and regulation of a GDNF-GFRα1 synaptic adhesion assembly.

Nature communications·2023
Same author

Decreased neonatal morbidity in 'stomach-down' left congenital diaphragmatic hernia: implications of prenatal ultrasound diagnosis for counseling and postnatal management.

Ultrasound in obstetrics & gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology·2021
Same author

Variability of nonpathogenic influenza virus H5N3 under immune pressure.

Acta virologica·2020
Same author

The impact of obesity on perceived patient-centred communication.

Obesity science & practice·2018
Same author

The Effectiveness of a Family-Centered Childhood Obesity Intervention at the YMCA: A Pilot Study.

Journal of community medicine & health education·2018
Same author

Conformationally restricted calpain inhibitors.

Chemical science·2017
Same journal

Lysozyme assay using a rationally designed GN4G2 substrate with coupled β-glucosidase reaction.

Analytical biochemistry·2026
Same journal

The long run: A tribute to Arthur Joseph Lawrence Cooper.

Analytical biochemistry·2026
Same journal

Evaluation of a method for affinity measurement using solution equilibrium titration with magnetic beads.

Analytical biochemistry·2026
Same journal

Metabolomics approach using UHPLC/QE-MS for the mechanism of He Xue Ming Mu tablets on non-proliferative diabetic retinopathy.

Analytical biochemistry·2026
Same journal

UniRES-GO: Unified residue-level early fusion of sequence and predicted structure for protein function prediction.

Analytical biochemistry·2026
Same journal

IgG detection by enzyme-linked mass spectrometric assay versus color, fluorescent, ECL in buffer and serum.

Analytical biochemistry·2026
See all related articles

We developed a fluorescence polarization assay to screen for inhibitors of the Src-SH2 domain, crucial for signaling pathways in diseases like cancer. This assay is robust and suitable for high-throughput screening of drug compounds.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • The tyrosine kinase pp60c-src is a therapeutic target for diseases such as cancer and osteoporosis.
  • The Src homology 2 (SH2) domain binds to phosphorylated tyrosine residues, regulating critical signaling pathways.
  • Inhibiting SH2 domain interactions offers potential therapeutic benefits for various human diseases.

Purpose of the Study:

  • To develop a versatile and reproducible assay for measuring compound binding to the Src-SH2 domain.
  • To facilitate high-throughput screening (HTS) of potential drug leads targeting SH2 interactions.

Main Methods:

  • A fluorescence polarization (FP) assay was designed to quantify the binding of small molecules to the Src-SH2 domain.
  • The assay demonstrates robustness, tolerating moderate optical density and up to 20% dimethyl sulfoxide (DMSO).

Related Experiment Videos

Main Results:

  • The developed FP assay is insensitive to fluorescence intensity variations.
  • The assay is suitable for screening large libraries of synthetic and natural compounds.
  • This method enables efficient identification of compounds that modulate SH2 domain interactions.

Conclusions:

  • The described fluorescence polarization assay is a valuable tool for high-throughput screening of Src-SH2 domain inhibitors.
  • This assay can accelerate the discovery of novel therapeutics targeting signaling pathways implicated in cancer and other diseases.