Analysis of DNA-Protein Interactions Using PAGE: Band-Shift Assays
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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 wordsBand shift DNA–protein interaction DNA binding constant EMSA PAGE Gel retardation
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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