A simple method for improving the specificity of anti-methyl histone antibodies.
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).
Connor
Caroline
C
Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts Medical School, Worcester, MA, USA.
Cheung
Iris
I
Simon
Andrew
A
Jakovcevski
Mira
M
Weng
Zhiping
Z
Akbarian
Schahram
S
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D006657:Histones / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D008745:Methylation; D024363:Transcription Initiation Site
D000906:Antibodies; D002843:Chromatin; D006657:Histones