Preparation of Site-Specific Antibodies to Acetylated Histones

Synthetic peptides corresponding to regions within the amino-terminal domains of the core histones H2A, H2B, H3, and H4, in which ϵ-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 ac...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 1999-11, Vol.19 (3), p.417-424
Main Authors: White, Darren A, Belyaev, Nikolai D, Turner, Bryan M
Format: Article
Language:English
Subjects:
Acetylation
Animals
Antibody Specificity
Histones - immunology
Histones - metabolism
Immunoglobulin G - isolation & purification
Immunologic Techniques
Rabbits
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Summary:Synthetic peptides corresponding to regions within the amino-terminal domains of the core histones H2A, H2B, H3, and H4, in which ϵ-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.
ISSN:1046-2023
1095-9130
DOI:10.1006/meth.1999.0878