The detection of DNA-binding proteins by protein blotting.
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.
D002352:Carrier Proteins / Q000032:analysis; D002850:Chromatography, Gel / Q000295:instrumentation / Q000379:methods; D004265:DNA Helicases / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006367:HeLa Cells / Q000032:analysis; D006657:Histones; D006801:Humans; D009698:Nucleoproteins / Q000032:analysis; D012367:RNA, Viral
D002352:Carrier Proteins; D006657:Histones; D009698:Nucleoproteins; D012367:RNA, Viral; D004265:DNA Helicases