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Analysis of DNA-Protein Interactions Using PAGE: Band-Shift Assays

  • Lynn Powell
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1054)

Abstract

The band-shift assay using polyacrylamide gel electrophoresis is a powerful technique used to investigate DNA–protein interactions. The basis of the method is the separation of free DNA from DNA–protein complexes by virtue of differences in charge, size, and shape. The band-shift assay can be used to determine thermodynamic and kinetic binding constants and also to analyze the composition and stoichiometries of DNA–protein complexes.

Key words

Band shift DNA–protein interaction DNA binding constant EMSA PAGE Gel retardation 

Notes

Acknowledgements

I would like to acknowledge the help of Professors Noreen Murray and Andrew Jarman in whose laboratories this work took place. Thanks are also due to my colleagues in the Murray and Jarman laboratories, in particular David Dryden, Laurie Cooper, and Petra zur Lage for their help and advice with protein purification.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Lynn Powell
    • 1
  1. 1.Centre for Integrative Physiology, School of Biomedical SciencesUniversity of EdinburghEdinburghUK

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