Nucleic Acids Research, 1980, Vol. 8, No. 1 1-20
© 1980
Articles |
The detection of DNA-binding proteins by protein blotting
+Department of Biochemical Sciences, Princeton University Princeton, NJ 08544 ++Department of Genetics, Hutchinson Cancer Center Seattle, WA 98104, USA
Received November 30, 1979. 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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