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From spots to beads— PTM ‐peptide bead arrays for the characterization of anti‐histone antibodies
Author(s) -
Heubach Yvonne,
Planatscher Hannes,
Sommersdorf Cornelia,
Maisch Daniel,
Maier Julia,
Joos Thomas O.,
Templin Markus F.,
Poetz Oliver
Publication year - 2013
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201200383
Subject(s) - immunoprecipitation , chromatin immunoprecipitation , histone , peptide , epigenetics , antibody , acetylation , bead , blot , histone h3 , chromatin , microbiology and biotechnology , computational biology , chemistry , biology , biochemistry , genetics , gene , gene expression , materials science , promoter , composite material
Antibodies that recognize PTMs of histones play a central role in epigenetic proteomic research. Modification‐specific antibodies are employed in chromatin immunoprecipitation, for W estern 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.