A new analytical scale DNA affinity binding assay for analyses of specific protein-DNA interactions

Anal Biochem. 2001 Dec 15;299(2):203-10. doi: 10.1006/abio.2001.5433.

Abstract

We describe a rapid analytical assay for identification of proteins binding to specific DNA sequences. The DAPSTER assay (DNA affinity preincubation specificity test of recognition assay) is a DNA affinity chromatography-based microassay that can discriminate between specific and nonspecific protein-DNA interactions. The assay is sensitive and can detect protein-DNA interactions and larger multicomponent complexes that can be missed by other analytical methods. Here we describe in detail the optimization and utilization of the DAPSTER assay to isolate AP-1 complexes and associated proteins in multimeric complexes bound to the AP-1 DNA element.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Binding Sites
  • Chromatography, Affinity / methods*
  • DNA / analysis*
  • DNA-Binding Proteins / analysis*
  • Egg Proteins / metabolism
  • HT29 Cells
  • Humans
  • Membrane Glycoproteins / metabolism
  • Oncogene Protein p65(gag-jun) / metabolism
  • Receptors, Cell Surface*
  • Transcription Factor AP-1 / metabolism
  • Tumor Cells, Cultured
  • Zona Pellucida Glycoproteins

Substances

  • DNA-Binding Proteins
  • Egg Proteins
  • Membrane Glycoproteins
  • Oncogene Protein p65(gag-jun)
  • Receptors, Cell Surface
  • Transcription Factor AP-1
  • Zona Pellucida Glycoproteins
  • DNA