From spots to beads-PTM-peptide bead arrays for the characterization of anti-histone antibodies

被引：10

作者：

Heubach, Yvonne ^{[1
]}

Planatscher, Hannes ^{[1
]}

Sommersdorf, Cornelia ^{[1
]}

Maisch, Daniel ^{[2
]}

Maier, Julia ^{[3
]}

Joos, Thomas O. ^{[1
]}

Templin, Markus F. ^{[1
]}

Poetz, Oliver ^{[1
]}

机构：

[1] Univ Tubingen, NMI Nat & Med Inst, D-72770 Reutlingen, Germany

[2] INTAVIS Bioanalyt Instruments AG, Heidelberg, Germany

[3] Univ Tubingen, Dept Pharmacol, Tubingen, Germany

来源：

PROTEOMICS | 2013年 / 13卷 / 06期

关键词：

Antibody characterisation; Epigenetics; Histone H3; Peptide bead array; Spot synthesis; Technology; MODIFIED HISTONE TAILS; SIGNALING NETWORKS; IMMUNOPRECIPITATION; SPECIFICITY; MICROARRAYS; PROTEIN; CELLS; ACETYLATION; AFFINITY; BINDING;

D O I：

10.1002/pmic.201200383

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：1010 / 1015

页数：6