Rapid, preparative-scale purification of chromatin proteins.
Methods are desceibed which permit rapid isolation of chromatographically purified histone and non-histone chromatin proteins under relatively mild chemical conditions. Chromatin is isolated from purified nuclei, dissociated in guanidine - HCl-urea and the nucleic acids removed by ultracentrigugation. This can be accomplished in 10 h by employing maximum-force rotors (500 000 x g). The proteins are then fractionated by a batch ion-exchange method, which leads to a rapid and complete separation of the histones and non-histone components, in apparently undegraded form. With these methods it is possible to obtain mg quantities of chromatographically pure histone and non-histone proteins in less than a single working day.
D001665:Binding Sites; D002843:Chromatin / Q000032:analysis*; D004273:DNA, Neoplasm / Q000032:analysis; D006146:Guanidines; D006367:HeLa Cells / Q000032:analysis; D006657:Histones / Q000302:isolation & purification; D009363:Neoplasm Proteins / Q000032:analysis; D009698:Nucleoproteins / Q000032:analysis / Q000302:isolation & purification*; D011485:Protein Binding; D012334:RNA, Neoplasm / Q000032:analysis; D014508:Urea
D002843:Chromatin; D004273:DNA, Neoplasm; D006146:Guanidines; D006657:Histones; D009363:Neoplasm Proteins; D009698:Nucleoproteins; D012334:RNA, Neoplasm; D014508:Urea