Determination of Histone H2AX Phosphorylation in DT40 Cells

  • Kana Nishihara
  • Sampada A. Shahane
  • Menghang XiaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1473)


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.

Key words

γH2AX High-contents imaging DSB Immunostaining 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Kana Nishihara
    • 1
  • Sampada A. Shahane
    • 1
  • Menghang Xia
    • 1
    Email author
  1. 1.National Center for Advancing Translational SciencesNational Institutes of HealthBethesdaUSA

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