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Cytometric Assessment Of Histone H2AX Phosphorylation: A Reporter Of DNA Damage.
X. Huang, Z. Darzynkiewicz
Published 2006 · Biology, Medicine
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DNA damage that leads to formation of DNA double-strand breaks (DSBs) induces phosphorylation of histone H2AX on Ser-139 at sites flanking the breakage. Immunocytochemical detection of phosphorylated H2AX (denoted as gammaH2AX) thus provides a marker of DSBs. The method presented in this chapter describes the detection of gammaH2AX for revealing the presence of DSBs, combined with differential staining of cellular DNA for revealing the cell cycle phase. The detection of gammaH2AX is based on indirect immunofluorescence using secondary antibody tagged with fluorescein isothiocyanate (FITC) while DNA is counterstained with propidium iodide (PI). Intensity of cellular green (FITC) and red (PI) fluorescence is measured by flow cytometry and bivariate analysis of the data is used to correlate the presence of DSBs with the cell cycle phase.
This paper references
DNA Double-stranded Breaks Induce Histone H2AX Phosphorylation on Serine 139*
E. Rogakou (1998)
Histone mRNA expression: multiple levels of cell cycle regulation and important developmental consequences.
W. Marzluff (2002)
Cell Cycle-Dependent Expression of Phosphorylated Histone H2AX: Reduced Expression in Unirradiated but not X-Irradiated G1-Phase Cells
S. H. Macphail (2003)
DNA-PK is activated by nucleosomes and phosphorylates H2AX within the nucleosomes in an acetylation-dependent manner.
Eunjung Park (2003)
DNA-damage signaling and apoptosis
N. Lowndes (2001)
Quantitative Detection of 125IdU-Induced DNA Double-Strand Breaks with γ-H2AX Antibody
O. Sedelnikova (2002)
ATM Phosphorylates Histone H2AX in Response to DNA Double-strand Breaks*
S. Burma (2001)
Assessment of histone H2AX phosphorylation induced by DNA topoisomerase I and II inhibitors topotecan and mitoxantrone and by the DNA cross‐linking agent cisplatin
X. Huang (2004)
Expression and radiation-induced phosphorylation of histone H2AX in mammalian cells.
K. Yoshida (2003)
Histone 2A, a heteromorphous family of eight protein species.
M. H. West (1980)
A role for Saccharomyces cerevisiae histone H2A in DNA repair
J. Downs (2000)
DNA Damage Induced by DNA Topoisomerase I- and Topoisomerase II- Inhibitors Detected by Histone H2AXphosphorylation in Relation to the Cell Cycle Phase and Apoptosis
X. Huang (2003)
Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo
E. Rogakou (1999)
Phosphorylation of Histone H2AX and Activation of Mre11, Rad50, and Nbs1 in Response to Replication-dependent DNA Double-strand Breaks Induced by Mammalian DNA Topoisomerase I Cleavage Complexes*
T. Furuta (2003)
Expression of phosphorylated histone H2AX in cultured cell lines following exposure to X‐rays
S. H. Macphail (2003)
DNA strand breaks occurring during apoptosis - their early insitu detection by the terminal deoxynucleotidyl transferase and nick translation assays and prevention by serine protease inhibitors.
W. Gorczyca (1992)
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