A simple method for improving the specificity of anti-methyl histone antibodies

Epigenetics. 2010 Jul 1;5(5):392-5. doi: 10.4161/epi.5.5.11874. Epub 2010 Jul 1.

Abstract

Antibodies differentiating between the mono-, di- and trimethylated forms of specific histone lysine residues are a critical tool in epigenome research, but show variable specificity, potentially limiting comparisons across studies and between samples. Using trimethyl histone H3 lysine 4 (H3K4me3)-a mark enriched at transcription start sites (TSS) of active genes-as an example, we describe how simple co-incubation with synthetic peptide of the K4me2 modification leads to increased specificity for K4me3 and a much sharper peak distribution proximal to TSS following chromatin immunoprecipitation and massively parallel sequencing (ChIP-Seq).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Antibodies / immunology
  • Antibody Specificity*
  • Cells, Cultured
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation / methods*
  • Histones / chemistry
  • Histones / immunology*
  • Histones / metabolism
  • Methylation
  • Transcription Initiation Site

Substances

  • Antibodies
  • Chromatin
  • Histones