The detection of DNA-binding proteins by protein blotting (original) (raw)

Journal Article

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Department of Biochemical Sciences, Princeton University

Princeton, NJ 08544

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Department of Biochemical Sciences, Princeton University

Princeton, NJ 08544

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Department of Biochemical Sciences, Princeton University

Princeton, NJ 08544

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Department of Genetics, Hutchinson Cancer Center

Seattle, WA 98104, USA

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Received:

30 November 1979

Published:

11 January 1980

Cite

Brian Bowen, Jay Steinberg, U.K. Laemmli, Harold Weintraub, The detection of DNA-binding proteins by protein blotting, Nucleic Acids Research, Volume 8, Issue 1, 11 January 1980, Pages 1–20, https://doi.org/10.1093/nar/8.1.1
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Abstract

A method, called “protein blotting,” for the detection of DNA-binding proteins is described. Proteins are separated on an SDS-polyacrylamide gel. The gel ia sandwiched between 2 nitrocellulose filters and the proteins allowed to diffuse out of the gel and onto the filters. The proteins are tightly bound to each filter, producing a replica of the original gel pattern. The replicais used to detect DNA-binding proteins, RKA-binding proteins or histonebinding proteins by incubation of the filter with [32P]DNA, [125I]RNA, or [125I] histone. Evidence is also presented that specific protein-DNA interactions may be detected by this technique; under appropriate conditions, the lac repressor binds only to DNA containing the lac operator. Strategies for the detection of specific protein-DNA interactions are discussed.

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