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Staining Against Phospho-H2AX (γ-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells

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Tumor Microenvironment

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 899))

Abstract

Phospho-H2AX or γ-H2AX- is a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalin-fixed, paraffin-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels.

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Author information

Authors and Affiliations

  1. Department of Molecular Materials, Institute for Molecules and Materials, Radboud University, Nijmegen, The Netherlands

    Anika Nagelkerke

  2. Department of Radiation Oncology, RadboudUMC, Nijmegen, The Netherlands

    Paul N. Span

Authors
  1. Anika Nagelkerke
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  2. Paul N. Span
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Corresponding author

Correspondence to Paul N. Span .

Editor information

Editors and Affiliations

  1. Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Constantinos Koumenis

  2. Cell & Developmental Biology, Oregon Health & Science University, Portland, Oregon, USA

    Lisa M. Coussens

  3. Department of Radiation Oncology, Stanford University School of Medic, Stanford, California, USA

    Amato Giaccia

  4. Department of Oncology, University of Oxford, Oxford, Oxfordshire, United Kingdom

    Ester Hammond

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© 2016 Springer International Publishing Switzerland

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Nagelkerke, A., Span, P.N. (2016). Staining Against Phospho-H2AX (γ-H2AX) as a Marker for DNA Damage and Genomic Instability in Cancer Tissues and Cells. In: Koumenis, C., Coussens, L., Giaccia, A., Hammond, E. (eds) Tumor Microenvironment. Advances in Experimental Medicine and Biology, vol 899. Springer, Cham. https://doi.org/10.1007/978-3-319-26666-4_1

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