Separation of nucleic acids and chromatin proteins by hydrophobic interaction chromatography.
A procedure by which chromatin proteins (histones and non-histones) can be rapidly separated from nucleic acids by hydrophobic interaction chromatography is described. The procedure is carried out under non-rigorous conditions that must be assumed to induce little irreversible change in the biological properties of most proteins. More than 90% (w/w) of the chromatin proteins can be retained by hydrophobic interaction while nucleic acids pass quantitatively through the columns. By gradient elution of the columns the histones can be divided into fractions containing H1, H2A/H2B and H3/H4, and at the same time a subfractionation of the non-histone proteins is obtained. Protein recovery depends on the type of column used, but exceeds 80% (w/w) with even the most strongly binding hydrophobic matrix investigated.
D000818:Animals; D002467:Cell Nucleus / Q000032:analysis; D002843:Chromatin / Q000032:analysis*; D002850:Chromatography, Gel / Q000379:methods; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D004247:DNA / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000302:isolation & purification*; D008099:Liver / Q000032:analysis; D008114:Liver Neoplasms, Experimental / Q000032:analysis; D008297:Male; D012313:RNA / Q000302:isolation & purification*; D051381:Rats; D011919:Rats, Inbred Strains
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D012313:RNA; D004247:DNA