title: Affinity reagents for studying histone modifications & guidelines for their quality control.
abstract: Histone post-translational modifications (PTMs) have pivotal functions in many chromatin processes, which makes their detection and characterization an imperative in chromatin biology. The established approaches for histone PTM characterization are generally based on affinity reagents specific for modified histone tails such as antibodies and, most recently, recombinant reading domains. Hence, the proper performance of these reagents is a critical precondition for the validity of the generated experimental data. In this review, we evaluate and update the quality criteria for assessment of the binding specificity of histone PTM affinity reagents. In addition, we discuss in detail the advantages and pitfalls of using antibodies and recombinant reading domains in chromatin biology research. Reading domains provide key advantages, such as consistent quality and recombinant production, but the future will tell if this emerging technology keeps its promises.
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, 70569 Stuttgart, Germany.
authors.forename: Goran
authors.initials: G
authors.lastname: Kungulovski
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, 70569 Stuttgart, Germany.
authors.forename: Rebekka
authors.initials: R
authors.lastname: Mauser
authors.affiliation: Institute of Biochemistry, Stuttgart University, Pfaffenwaldring 55, 70569 Stuttgart, Germany.
authors.forename: Albert
authors.initials: A
authors.lastname: Jeltsch
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D000915:Antibody Affinity; D015153:Blotting, Western / Q000379:methods; D002843:Chromatin / Q000737:chemistry* / Q000378:metabolism; D047369:Chromatin Immunoprecipitation / Q000379:methods; D015894:Genome, Human; D017408:Guidelines as Topic; D006657:Histones / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D040081:Protein Array Analysis / Q000379:methods; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011786:Quality Control; D011994:Recombinant Proteins / Q000737:chemistry*
chemical_list: D000906:Antibodies; D002843:Chromatin; D006657:Histones; D011994:Recombinant Proteins
keywords: ChIP; antibodies; histone modifications; nucleosome; quality control; reading domains; specificity