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Phosphorylated H2AX (γ-H2AX) is a sensitive marker for DNA double-strand breaks (DSBs), but the variability of H2AX expression in different cell and tissue types makes it difficult to interpret the meaning of the γ-H2AX level. Furthermore, the assays commonly used for γ-H2AX detection utilize laborious and low-throughput microscopy-based methods. We describe here an ELISA assay that measures both phosphorylated H2AX and total H2AX absolute amounts to determine the percentage of γ-H2AX, providing a normalized value representative of the amount of DNA damage. We demonstrate the utility of the assay to measure DSBs introduced by either ionizing radiation or DNA-damaging agents in cultured cells and in xenograft models. Furthermore, utilizing the NCI-60 cancer cell line panel, we show a correlation between the basal fraction of γ-H2AX and cellular mutation levels. This additional application highlights the ability of the assay to measure γ-H2AX levels in many extracts at once, making it possible to correlate findings with other cellular characteristics. Overall, the γ-H2AX ELISA represents a novel approach to quantifying DNA damage, which may lead to a better understanding of mutagenic pathways in cancer and provide a useful biomarker for monitoring the effectiveness of DNA-damaging anticancer agents. [ABSTRACT FROM AUTHOR]

Theoretical predictions suggest the presence of tetrahedral symmetry as an explanation for the vanishing intra-band E2 transitions at the bottom of the odd-spin negative-parity band in 156Gd. The present study reports on experiment performed to address this phenomenon. It allowed to remove certain ambiguouities related to the intra-band E2 transitions in the negative-parity bands, to determine the new inter-band transitions and reduced probability ratios B(E2)/B(E1) and, for the first time, to determine the experimental uncertainties related to the latter observable. [ABSTRACT FROM AUTHOR]