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

Anika Nagelkerke, Paul N Span

Research output: Contribution to journalArticleAcademicpeer-review

49 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)1-10
Number of pages10
JournalAdvances in experimental medicine and biology
Publication statusPublished - 22-Jun-2016
Externally publishedYes


  • Biomarkers/metabolism
  • Cryoultramicrotomy
  • DNA Damage
  • Genomic Instability
  • Histones/metabolism
  • Humans
  • Neoplasms/genetics
  • Paraffin Embedding
  • Phosphorylation
  • Staining and Labeling/methods
  • Tissue Fixation

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