An assessment of histone-modification antibody quality

Department of Molecular, Cell and Developmental Biology, University of California Santa Cruz, Santa Cruz, California, USA.
Nature Structural & Molecular Biology (Impact Factor: 13.31). 01/2011; 18(1):91-3. DOI: 10.1038/nsmb.1972
Source: PubMed


We have tested the specificity and utility of more than 200 antibodies raised against 57 different histone modifications in Drosophila melanogaster, Caenorhabditis elegans and human cells. Although most antibodies performed well, more than 25% failed specificity tests by dot blot or western blot. Among specific antibodies, more than 20% failed in chromatin immunoprecipitation experiments. We advise rigorous testing of histone-modification antibodies before use, and we provide a website for posting new test results (

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    • "To this aim, we have developed a collection of native and engineered histone modification interaction domains (HMIDs), which strongly and specifically interact with H3 tails methylated at different lysine residues. The most promising domains were investigated with respect to the primary and secondary ENCODE criteria for antibody validation, such as specificity in Western blotting with native, recombinant, and PTM-depleted histones, binding to modified peptides, and ChIP-like experiments (Egelhofer et al. 2011; Landt et al. 2012). In all our experiments, the efficiency of HMIDs was directly compared side-by-side with popular commercial antibodies recognizing the same PTMs. "
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