The detection of DNA-binding proteins by protein blotting

Brian C. Bowen; Jay A. Steinberg; Ulrich K. Laemmli; Harold Weintraub

doi:10.1093/nar/8.1.1

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

BiologyDNAMolecular biologyDNA-binding proteinBlotGel electrophoresisNon-histone proteinBiochemistryRepressorHistoneGeneGene expressionTranscription factor

Affiliated Institutions

Princeton University US

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Publication Info

Year: 1980
Type: article
Volume: 8
Issue: 1
Pages: 1-20
Citations: 731
Access: Closed

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APA Style

                            
                                
                                    Brian C. Bowen, 
                                
                                    Jay A. Steinberg, 
                                
                                    Ulrich K. Laemmli
                                
                                et al.
                            
                            (1980). 
                            The detection of DNA-binding proteins by protein blotting. 
                            Nucleic Acids Research
                            , 8
                            (1)
                            , 1-20.
                            https://doi.org/10.1093/nar/8.1.1
                        

Identifiers

DOI: 10.1093/nar/8.1.1