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Detection of DNA Double-Strand Breaks by γ-H2AX Immunodetection

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Homologous Recombination

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2153))

Abstract

DNA double-strand breaks (DSBs) are the most deleterious type of DNA damage and a cause of genetic instability as they can lead to mutations, genome rearrangements, or loss of genetic material when not properly repaired. Eukaryotes from budding yeast to mammalian cells respond to the formation of DSBs with the immediate phosphorylation of a histone H2A isoform. The modified histone, phosphorylated in serine 139 in mammals (S129 in yeast), is named γ-H2AX. Detection of DSBs is of high relevance in research on DNA repair, aging, tumorigenesis, and cancer drug development, given the tight association of DSBs with different diseases and its potential to kill cells. DSB levels can be obtained by measuring levels of γ-H2AX in extracts of cell populations or by counting foci in individual nuclei. In this chapter some techniques to detect γ-H2AX are described.

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Acknowledgments

AA’s lab is funded by the Spanish Ministry of Economy and Competitiveness, Junta de Andalucía, European Research Council, and the European Union (FEDER).

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Authors and Affiliations

  1. Centro Andaluz de Biología Molecular y Medicina Regenerativa, CABIMER, University of Seville-CSIC-UPO, Seville, Spain

    Sonia I. Barroso & Andrés Aguilera

Authors
  1. Sonia I. Barroso
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  2. Andrés Aguilera
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Corresponding author

Correspondence to Andrés Aguilera .

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Editors and Affiliations

  1. Centro Andaluz de Biologı´a Molecular, y Medicina Regenerativa, CABIMER, Universidad de Sevilla, Sevilla, Spain

    Andrés Aguilera

  2. Institut Curie, PSL Research University, CNRS, UMR3348, F-91405, Orsay, France

    Aura Carreira

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Barroso, S.I., Aguilera, A. (2021). Detection of DNA Double-Strand Breaks by γ-H2AX Immunodetection. In: Aguilera, A., Carreira, A. (eds) Homologous Recombination. Methods in Molecular Biology, vol 2153. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0644-5_1

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  • DOI: https://doi.org/10.1007/978-1-0716-0644-5_1

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0643-8

  • Online ISBN: 978-1-0716-0644-5

  • eBook Packages: Springer Protocols

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