title: From spots to beads-PTM-peptide bead arrays for the characterization of anti-histone antibodies.
abstract: Antibodies that recognize PTMs of histones play a central role in epigenetic proteomic research. Modification-specific antibodies are employed in chromatin immunoprecipitation, for Western blotting and during the immunoprecipitation steps for MS-based global proteomic analyses. Knowledge about the antibodies' off-target binding is essential for the interpretation of experimental data. To address this challenge we developed a fast and cost efficient system for generating peptide bead arrays. We employed this method to establish a bead-based peptide array containing 384 peptides displaying phosphorylated, acetylated, methylated, and citrullinated N-terminal regions of histones H2A, H2B, H3 and H4 and controls. We profiled the binding of 40 PTM-specific antibodies important for epigenetic proteomic research.
authors: NMI Natural and Medical Sciences Institute at the University of Tuebingen, Reutlingen, Germany.
authors: Yvonne
authors: Y
authors: Heubach
authors: Hannes
authors: H
authors: Planatscher
authors: Cornelia
authors: C
authors: Sommersdorf
authors: Daniel
authors: D
authors: Maisch
authors: Julia
authors: J
authors: Maier
authors: Thomas O
authors: TO
authors: Joos
authors: Markus F
authors: MF
authors: Templin
authors: Oliver
authors: O
authors: Poetz
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D044127:Epigenesis, Genetic; D000939:Epitopes / Q000737:chemistry; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D010455:Peptides; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D040901:Proteomics
chemical_list: D000906:Antibodies; D000939:Epitopes; D006657:Histones; D010455:Peptides