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Electrophoretic assay for DNA-binding proteins

Varshavsky, Alexander (1987) Electrophoretic assay for DNA-binding proteins. In: Methods in Enzymology. Vol.151. Academic Press , pp. 551-565. ISBN 9780121820527. https://resolver.caltech.edu/CaltechAUTHORS:20210122-162658053

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Abstract

This chapter describes a generally applicable assay for specific DNA-binding proteins in crude extracts. The assay is based on gel electrophoretic separation of protein–DNA complexes from each other and from free DNA. This assay employed in several laboratories to detect a variety of DNA-binding proteins. In spite of relatively brief experience with the electrophoretic assay as a detection tool, its exceptionally high sensitivity has already made this assay the method of choice in many experimental settings. The electrophoretic assay has been successfully applied to the detection of both highly sequence-specific, nonabundant DNA-binding proteins and abundant DNA-binding proteins of relatively low nucleotide sequence specificity. After briefly describing some of the alternative techniques suitable for the detection of DNA-binding proteins in crude extracts, it discusses the electrophoretic assay, its recent modifications, and consider other applications of the assay. Although nondenaturing electrophoretic systems have existed for almost as long as the method of gel electrophoresis itself, the arrival of powerful analytical applications of this technology is quite recent and other useful variations of the theme are still to come.


Item Type:Book Section
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https://doi.org/10.1016/s0076-6879(87)51044-8DOIArticle
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Varshavsky, Alexander0000-0002-4011-258X
Additional Information:© 1987 by Academic Press. I thank my colleagues at MIT and elsewhere for permission to reproduce their published results in Figs. 3 and 4, and for providing preprints describing their unpublished work. I also thank Daniel Finley, Peter Kolodziej, Edward Winter, and especially Mark Solomon for their comments on the manuscript. Work in the author's laboratory is supported by grants from the National Institutes of Health.
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Record Number:CaltechAUTHORS:20210122-162658053
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20210122-162658053
Official Citation:Alexander Varshavsky, Electrophoretic assay for DNA-binding proteins, Methods in Enzymology, Academic Press, Volume 151, 1987, Pages 551-565, ISSN 0076-6879, ISBN 9780121820527, https://doi.org/10.1016/S0076-6879(87)51044-8. (http://www.sciencedirect.com/science/article/pii/S0076687987510448)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:107683
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:25 Jan 2021 15:08
Last Modified:25 Jan 2021 15:08

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