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Methods Mol Biol. 2016;1473:71-6. doi: 10.1007/978-1-4939-6346-1_8.

Determination of Histone H2AX Phosphorylation in DT40 Cells.

Author information

1
National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA.
2
National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA. mxia@mail.nih.gov.

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.

KEYWORDS:

DSB; High-contents imaging; Immunostaining; γH2AX

PMID:
27518625
PMCID:
PMC5375169
DOI:
10.1007/978-1-4939-6346-1_8
[Indexed for MEDLINE]
Free PMC Article

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