Preparation of site-specific antibodies to acetylated histones

Abstract

Synthetic peptides corresponding to regions within the amino-terminal domains of the core histones H2A, H2B, H3, and H4, in which epsilon-acetyllysine has been substituted for selected lysines, have been used to raise polyclonal antisera in rabbits. Such antisera can be specific not only for individual acetylated histones but also for histone isoforms acetylated at particular lysine residues. In this article, we describe procedures for the preparation, affinity purification, and initial characterization of site-specific antisera to acetylated histones.