Literature DB >> 16673875

Cytometric assessment of histone H2AX phosphorylation: a reporter of DNA damage.

Xuan Huang1, Zbigniew Darzynkiewicz.   

Abstract

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.

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Year:  2006        PMID： 16673875      PMCID： PMC1458374          DOI： 10.1385/1-59259-973-7:073

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN： 1064-3745


  18 in total

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