title: The development of a monoclonal antibody recognizing the Drosophila melanogaster phosphorylated histone H2A variant (γ-H2AV).
abstract: The recognition of DNA double-strand breaks (DSBs) using a phospho-specific antibody to the histone 2A variant has become the gold standard assay for DNA damage detection. Here we report on the development of the first monoclonal antibody to the phospho-specific form of Drosophila H2AV and characterize the specificity of this antibody to programmed DSBs in oocytes and rereplication sites in endocycling cells by immunofluorescence assays and to DSBs resulting from irradiation in both cell culture and whole tissue by Western blot assays. These studies show that the antibody derived in the study is highly specific for this modification that occurs at DSB sites, and therefore will be a new useful tool within the Drosophila community for the study of DNA damage response, DSB repair, meiotic recombination and chemical agents that cause DNA damage.
authors.affiliation: Stowers Institute for Medical Research, Kansas City, Missouri 64110.
authors.forename: Cathleen M
authors.initials: CM
authors.lastname: Lake
authors.forename: Julie Korda
authors.initials: JK
authors.lastname: Holsclaw
authors.forename: Stephanie P
authors.initials: SP
authors.lastname: Bellendir
authors.forename: Jeff
authors.initials: J
authors.lastname: Sekelsky
authors.forename: R Scott
authors.initials: RS
authors.lastname: Hawley
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development; D019085:Fluorescent Antibody Technique, Direct; D006657:Histones / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D008540:Meiosis; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010766:Phosphorylation; D011839:Radiation, Ionizing
chemical_list: D000911:Antibodies, Monoclonal; D006657:Histones
keywords: DNA repair; H2AX; double-strand break; meiosis; γ-H2AV