title: Production and Purification of Antibodies Against Histone Modifications.
abstract: Antibodies that recognize specific histone modifications are invaluable tools to study chromatin structure and function. There are numerous commercially available antibodies that recognize a remarkable diversity of histone modifications. Unfortunately, many of them fail to work in certain applications or lack the high degree of specificity required of these reagents. The production of affinity-purified polyclonal antibodies against histone modifications demands a little effort but, in return, provides extremely valuable tools that overcome many of the concerns and limitations of commercial antibodies. We present a series of protocols and guidelines for the production and use of large amounts of polyclonal antibodies that recognize modifications of canonical histones. Our protocols can be applied to obtain antibodies that occur in histone variants and proteins other than histones. In addition, some of our protocols are compatible with the production of monoclonal or recombinant antibodies.
authors.affiliation: Department of Biology, Université de Sherbrooke, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Benoit
authors.initials: B
authors.lastname: Guillemette
authors.affiliation: Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, QC, Canada, H1T 2M4.
authors.forename: Ian
authors.initials: I
authors.lastname: Hammond-Martel
authors.affiliation: Centre de Recherche de l'Hôpital Maisonneuve-Rosemont, Montréal, QC, Canada, H1T 2M4.|Département de Médecine, Université de Montréal, Montréal, QC, Canada, H3T 1J4.
authors.forename: Hugo
authors.initials: H
authors.lastname: Wurtele
authors.affiliation: Institut de Recherche en Immunologie et Cancérologie (IRIC), Université de Montréal, CP 6128, Succursale Centre-Ville, Montréal, QC, Canada, H3C 3J7. alain.verreault@umontreal.ca.|Département de Pathologie et Biologie Cellulaire, Université de Montréal, CP 6128, Succursale Centre-Ville, Montréal, QC, Canada, H3C 3J7. alain.verreault@umontreal.ca.
authors.forename: Alain
authors.initials: A
authors.lastname: Verreault
mesh_terms: D000107:Acetylation; D000818:Animals; D002843:Chromatin / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D009707:Nucleosomes / Q000378:metabolism; D011499:Protein Processing, Post-Translational
chemical_list: D002843:Chromatin; D006657:Histones; D009707:Nucleosomes
keywords: Acetylation; Antibodies; Chromatin; Histone modification; Methylation; Nucleosome; Polyclonal