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Determination of Histone H2AX Phosphorylation in DT40 Cells

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High-Throughput Screening Assays in Toxicology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1473))

Abstract

Visualization of DNA damage response protein recruitment to DNA damage sites enables measurement of the DNA damage. DNA double-strand breaks (DSBs) and blocked replication forks induce the phosphorylation of H2AX at serine 139 (γH2AX), and accumulate γH2AX which can then be detected as foci. The detection of γH2AX foci by immunostaining with antibodies that recognize γH2AX is an indicator of DSBs presence. This chapter describes the measurement of γH2AX immunostaining using a high-content imaging platform in chicken DT40 B-lymphocyte cell lines.

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Authors and Affiliations

  1. National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA

    Kana Nishihara, Sampada A. Shahane & Menghang Xia

Authors
  1. Kana Nishihara
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  2. Sampada A. Shahane
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  3. Menghang Xia
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Corresponding author

Correspondence to Menghang Xia .

Editor information

Editors and Affiliations

  1. Department of Chemistry Rutgers Center for Computational and Integrative Biology, Rutgers University, Camden, New Jersey, USA

    Hao Zhu

  2. National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA

    Menghang Xia

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© 2016 Springer Science+Business Media New York

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Nishihara, K., Shahane, S.A., Xia, M. (2016). Determination of Histone H2AX Phosphorylation in DT40 Cells. In: Zhu, H., Xia, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 1473. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6346-1_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6346-1_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6344-7

  • Online ISBN: 978-1-4939-6346-1

  • eBook Packages: Springer Protocols

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