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An assessment of histone-modification antibody quality


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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Figure 1: Representative western assays and results.
Figure 2: Representative dot blot assays and results.
Figure 3: ChIP-chip and ChIP-seq.


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This work was supported by US National Institutes of Health (NIH) modENCODE grants U01HG004270 to J.D.L. and U01HG004258 to G.H.K., and by the Reference Epigenome Roadmap project grant U01ES017166 to B.R. We thank H. Kimura (Kyoto University) and T. Jenuwein (Max Planck Institute of Immunobiology) for providing the noncommercial antibodies indicated in Supplementary Table 1 at no cost.

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J.A., A.A.A., M.-S.C., D.S.D., T.A.E., S.C.R.E., S.G., A.A.G., T.G., R.D.H., G.H.K., P.V.K., S. Klugman, P.K.-Z., S. Kuan, M.I.K., I.L., K.L., J.D.L., D.L.-B., Y.L., A.M., Q.N., P.J.P., M.P., V.P., A.R., B.R., N.C.R., Y.B.S., G.A.S., S.S. and A.V. designed, executed, and analyzed the experiments. T.A.E., R.D.H., G.H.K., J.D.L., A.M. and S.S. wrote the manuscript.

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Correspondence to Peter J Park, Gary H Karpen, R David Hawkins or Jason D Lieb.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 (PDF 250 kb)

Supplementary Table 1

Tested antibodies and results. (XLS 130 kb)

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Egelhofer, T., Minoda, A., Klugman, S. et al. An assessment of histone-modification antibody quality. Nat Struct Mol Biol 18, 91–93 (2011).

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