A simple method for improving the specificity of anti-methyl histone antibodies
AuthorsConnor, Caroline M.
Student AuthorsCaroline Connor
UMass Chan AffiliationsBrudnick Neuropsychiatric Research Institute, Department of Psychiatry
Program in Bioinformatics and Integrative Biology
Department of Biochemistry and Molecular Pharmacology
KeywordsAmino Acid Sequence; Antibodies; *Antibody Specificity; Cells, Cultured; Chromatin; Chromatin Immunoprecipitation; Histones; Methylation; Transcription Initiation Site
Medicine and Health Sciences
Neuroscience and Neurobiology
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AbstractAntibodies 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).
SourceEpigenetics. 2010 Jul 1;5(5):392-5. Epub 2010 Jul 1. Link to article on publisher's website
Permanent Link to this Itemhttp://hdl.handle.net/20.500.14038/33150
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RightsThis is an open access article licensed under a under a Creative Commons Attribution-NonCommercial 3.0 Unported License.