title: Determination of Histone H2AX Phosphorylation in DT40 Cells.
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.
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA.
authors: Kana
authors: K
authors: Nishihara
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Building C, MSC: 3375, 9800 Medical Center Drive, Bethesda, MD, 20892, USA.
authors: Sampada A
authors: SA
authors: Shahane
authors: 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.
authors: Menghang
authors: M
authors: Xia
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D002645:Chickens; D004247:DNA / Q000235:genetics / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology*; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods*; D008558:Melphalan / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects
chemical_list: D000906:Antibodies; D000970:Antineoplastic Agents; D006657:Histones; D004317:Doxorubicin; D004247:DNA; D008558:Melphalan
keywords: DSB; High-contents imaging; Immunostaining; γH2AX