Validation of a cell-based screen for insulin receptor modulators by quantification of insulin receptor phosphorylation

J Biomol Screen. 2003 Aug;8(4):439-46. doi: 10.1177/1087057103255280.

Abstract

Stimulation of a cell with insulin initiates a signal transduction cascade that results in cellular activities that include phosphorylation of the receptor itself. Measurement of the degree of phosphorylation can serve as a marker for receptor activation. Receptor phosphorylation has been measured using Western blot analysis, which is very low throughput and not easily quantifiable. The goal of this project was to develop a cell-based assay to measure receptor phosphorylation in high throughput. This report describes a cell-based assay for insulin receptor phosphorylation that is robust and amenable to high-volume screening in a microwell format.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Fluorescence Resonance Energy Transfer / methods*
  • Indicators and Reagents
  • Insulin / metabolism*
  • Phosphorylation
  • Protein-Tyrosine Kinases / metabolism
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Receptor, Insulin / metabolism*
  • Signal Transduction

Substances

  • Indicators and Reagents
  • Insulin
  • Protein-Tyrosine Kinases
  • Receptor Protein-Tyrosine Kinases
  • Receptor, Insulin