title: Acid-Urea Gel Electrophoresis and Western Blotting of Histones.
abstract: Acid-urea gel electrophoresis offers significant advantages over SDS-PAGE for analysis of post-translational protein modifications, being capable of resolving proteins of similar size but varying in charge. Hence, it can be used to separate protein variants with small charge-altering differences in primary sequence, and is particularly useful in the analysis of histones whose charge variation arises from post-translational modification, such as phosphorylation or acetylation. On acid-urea gels, histones that carry multiple modifications, each with a characteristic charge, are resolved into distinct bands, the so-called "histone ladder." Thus, the extent and distribution of different modification states of histones can be visualized. Here, we describe the analysis of histone H3 by acid-urea gel electrophoresis and western blotting.
authors.affiliation: Nuclear Signalling Laboratory, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Hazzalin
authors.affiliation: Nuclear Signalling Laboratory, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK. louis.mahadevan@bioch.ox.ac.uk.
authors.forename: Louis C
authors.initials: LC
authors.lastname: Mahadevan
mesh_terms: D000107:Acetylation; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D004586:Electrophoresis / Q000379:methods*; D006657:Histones / Q000378:metabolism*; D051379:Mice; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D014508:Urea / Q000737:chemistry*
chemical_list: D006657:Histones; D014508:Urea
keywords: Acetylation; Acid-urea (AU) gel electrophoresis; Histone H3; Histone extraction; Histone ladder; Phosphorylation; Western blotting