Demonstrating Interactions of Transcription Factors with DNA by Electrophoretic Mobility Shift Assay

Methods Mol Biol. 2017:1651:11-21. doi: 10.1007/978-1-4939-7223-4_2.

Abstract

Confirming the binding of a transcription factor with a particular DNA sequence may be important in characterizing interactions with a synthetic promoter. Electrophoretic mobility shift assay is a powerful approach to demonstrate the specific DNA sequence that is bound by a transcription factor and also to confirm the specific transcription factor involved in the interaction. In this chapter we describe a method we have successfully used to demonstrate interactions of endogenous transcription factors with sequences derived from endogenous and synthetic promoters.

Keywords: EMSA; Electrophoretic mobility shift assay; Supershift; Transcription factor; Transcription factor binding site.

MeSH terms

  • Base Sequence
  • Binding Sites
  • Cell Line
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism*
  • Electrophoretic Mobility Shift Assay / methods*
  • Erythroid-Specific DNA-Binding Factors / metabolism
  • Humans
  • Jurkat Cells
  • NF-kappa B / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • Transcription Factors / metabolism*

Substances

  • Erythroid-Specific DNA-Binding Factors
  • NF-kappa B
  • Transcription Factors
  • DNA