Scientific article
OA Policy
English

The detection of DNA-binding proteins by protein blotting

Published inNucleic acids research, vol. 8, no. 1, p. 1-20
Publication date1980-01-11
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.

Keywords
  • Carrier Proteins / analysis
  • Chromatography, Gel / instrumentation
  • Chromatography, Gel / methods
  • DNA Helicases / analysis
  • Electrophoresis, Polyacrylamide Gel / methods
  • HeLa Cells / analysis
  • Histones
  • Humans
  • Nucleoproteins / analysis
  • RNA, Viral
Affiliation entities Not a UNIGE publication
Citation (ISO format)
BOWEN, Brian et al. The detection of DNA-binding proteins by protein blotting. In: Nucleic acids research, 1980, vol. 8, n° 1, p. 1–20. doi: 10.1093/nar/8.1.1
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Article (Published version)
Identifiers
Journal ISSN0305-1048
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312downloads

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