Fluorescence gel retardation assay to detect protein-protein interactions

Methods Mol Biol. 2004:261:155-60. doi: 10.1385/1-59259-762-9:155.

Abstract

A gel mobility retardation assay can be used to detect a protein-protein interaction. The assay is based on the electrophoretic mobility of a protein-protein complex being less than that of either protein alone. Electrophoretic mobility is detected by the fluorescence of a green fluorescent protein variant that is fused to one of the protein partners. The assay is demonstrated by using the interaction of the S-protein and S-peptide fragments of ribonuclease A as a case study.

Publication types

  • Review

MeSH terms

  • Animals
  • Electrophoretic Mobility Shift Assay / methods*
  • Fluorescence
  • Green Fluorescent Proteins
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Protein Binding
  • Proteins / chemistry*
  • Proteins / metabolism*

Substances

  • Luminescent Proteins
  • Proteins
  • Green Fluorescent Proteins