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An assessment of histone-modification antibody quality

Nature Structural & Molecular Biology volume 18, pages 91–93 (2011)Cite this article

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Abstract

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 (http://compbio.med.harvard.edu/antibodies/).

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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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Acknowledgements

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.

Author information

Author notes

  1. Thea A Egelhofer, Aki Minoda and Sarit Klugman: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular, Cell and Developmental Biology, University of California Santa Cruz, Santa Cruz, California, USA

    Thea A Egelhofer, Andreas Rechtsteiner & Susan Strome

  2. Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, California, USA

    Aki Minoda & Gary H Karpen

  3. Department of Genome Dynamics, Lawrence Berkeley National Lab, Berkeley, California, USA

    Aki Minoda & Gary H Karpen

  4. Ludwig Institute for Cancer Research, La Jolla, California, USA

    Sarit Klugman, Samantha Kuan, Ying Luu, Queminh Ngo, Bing Ren & R David Hawkins

  5. Department of Cellular and Molecular Medicine, University of California, San Diego School of Medicine, La Jolla, California, USA.,

    Sarit Klugman, Samantha Kuan, Ying Luu, Queminh Ngo, Bing Ren & R David Hawkins

  6. Center for Biomedical Informatics, Harvard Medical School, Boston, Massachusetts, USA

    Kyungjoon Lee, Daniel S Day, Peter V Kharchenko & Peter J Park

  7. The Gurdon Institute, University of Cambridge, Cambridge, UK

    Paulina Kolasinska-Zwierz, Ming-Sin Cheung, Isabel Latorre, Anne Vielle & Julie Ahringer

  8. Department of Genetics, University of Cambridge, Cambridge, UK

    Paulina Kolasinska-Zwierz, Ming-Sin Cheung, Isabel Latorre, Anne Vielle & Julie Ahringer

  9. Division of Genetics, Department of Medicine, Brigham & Women's Hospital, Boston, Massachusetts, USA

    Artyom A Alekseyenko, Andrey A Gorchakov & Mitzi I Kuroda

  10. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Artyom A Alekseyenko, Andrey A Gorchakov & Mitzi I Kuroda

  11. Department of Biology, Washington University in St. Louis, St. Louis, Missouri, USA

    Sarah Gadel, Tingting Gu, Nicole C Riddle & Sarah C R Elgin

  12. Department of Molecular Biology & Biochemistry, Rutgers University, Piscataway, New Jersey, USA

    Daniela Linder-Basso, Yuri B Schwartz, Gregory A Shanower & Vincenzo Pirrotta

  13. Ontario Institute for Cancer Research, Toronto, Ontario, Canada

    Marc Perry

  14. Department of Biology, Carolina Center for Genome Sciences, Chapel Hill, North Carolina, USA

    Jason D Lieb

  15. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA

    Jason D Lieb

Authors
  1. Thea A Egelhofer
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  2. Aki Minoda
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  3. Sarit Klugman
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  4. Kyungjoon Lee
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  5. Paulina Kolasinska-Zwierz
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  6. Artyom A Alekseyenko
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  8. Daniel S Day
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  9. Sarah Gadel
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  17. Queminh Ngo
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  18. Marc Perry
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  19. Andreas Rechtsteiner
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  20. Nicole C Riddle
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  21. Yuri B Schwartz
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  22. Gregory A Shanower
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  23. Anne Vielle
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  24. Julie Ahringer
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  25. Sarah C R Elgin
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  28. Bing Ren
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  29. Susan Strome
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  30. Peter J Park
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  31. Gary H Karpen
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  32. R David Hawkins
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  33. Jason D Lieb
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Contributions

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.

Corresponding authors

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

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Competing interests

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). https://doi.org/10.1038/nsmb.1972

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