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PMID: 6243775 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The detection of DNA-binding proteins by protein blotting.

Nucleic acids research ·Vol. 8 ·No. 1 ·1980-01-11 ·Pages 1-20

Bowen B, Steinberg J, Laemmli UK, Weintraub H

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.

MeSH Terms

Carrier Proteins/analysis Chromatography, Gel/instrumentation,methods DNA Helicases/analysis Electrophoresis, Polyacrylamide Gel/methods HeLa Cells/analysis Histones Humans Nucleoproteins/analysis RNA, Viral

Chemicals

Carrier Proteins Histones Nucleoproteins RNA, Viral DNA Helicases

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Bowen B

Steinberg J

Laemmli U K

Weintraub H

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26 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/8.1.1

Published

1980-01-11

Pages

1-20

Language

English

Region

England

NLM ID

0411011

PMCID

PMC327239

Subset

External Links

PubMed source DOI: 10.1093/nar/8.1.1 PMC Full Text View journal details

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