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. 1980 Jan 11;8(1):1-20.
doi: 10.1093/nar/8.1.1.

The detection of DNA-binding proteins by protein blotting

B Bowen, J Steinberg, U K Laemmli, H Weintraub

The detection of DNA-binding proteins by protein blotting

B Bowen et al. Nucleic Acids Res. .

Abstract

A method, called "protein blotting," for the detection of DNA-binding proteins is described. Proteins are separated on an SDA-polyacrylamide gel. The gel is 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 replica is used to detect DNA-binding proteins, RNA-binding proteins or histone-binding 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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