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There is a paucity of large animal models to study both the extent and the health risk of ionizing radiation exposure in humans. One promising candidate for such a model is the minipig. Here, we evaluate the minipig for its potential in γ-H2AX-based biodosimetry after exposure to ionizing radiation using both Cs137 and Co60 sources. γ-H2AX foci were enumerated in blood lymphocytes and normal Abstract Purpose: Within the EU RENEB project, seven laboratories have taken part in training and harmonisation activities to strengthen triage gamma-H2AX-based radiation exposure assessment. This has culminated in a second triage biodosimetry exercise.

Gamma h2ax assay

Abingdon, UK & Lake Forest CA, USA — AMSBIO announces the first commercially available gamma H2AX Pharmacodynamic assay kit for the study of double strand DNA breaks through the detection of gamma H2AX – a phosphorylated histone historically proven as a highly specific and sensitive molecular marker for double strand DNA damage detection. The gamma-H2AX assay has up to now not been applied in this context, and it is a promising tool for investigating the early cellular response to mixed beam irradiation. To determine the dose response and repair kinetics of gamma-H2AX ionizing radiation-induced foci in VH10 human fibroblasts exposed to mixed beams of 241Am alpha particles and X-rays. 2016-03-04 · The gammaH2AX assay for genotoxic and nongenotoxic agents: comparison of H2AX phosphorylation with cell death response. Toxicol Sci 140, 103–117 (2014). CAS Article Google Scholar A gamma-H2AX kit and antibody allows the assessment of DNA damage and DNA repair for ELISA based assays, immunohistochemistry or flow cytometry.

Immunostaining was then performed using the parameters listed in Table 1.

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Regarding case-control studies, as the main purpose of using H2AX assay in this case is to evaluate the persistent levels of phosphorylated histone as indicative of DNA damage already fixed (Sedelnikova et al., 2004), no previous stimulation is necessary to carry out the assay since the remaining γH2AX will be already present in the DNA without requiring cell division. In such studies, this approach is applied as a biomarker of genomic instability, likely as a result of deficiencies in DNA Gamma-H2AX is used widely as DNA damage marker in vitro, but its use for genotoxicity assessment in vivo has not been extensively investigated. Here, we developed an image analysis system for the precise quantification of the gamma-H2AX signal, which we used to monitor DNA damage in animals treated with known genotoxicants (EMS, ENU and doxorubicin). Laboratories around the world now use the gamma-H2AX assay developed by Bonner to study how cells sense and respond to double-strand breaks.

Assaying DNA Damage in Hippocampal Neurons Using the

Histone H2AX: Products. Histone H2AX is one of a number of core histone proteins. In the cellular response to genotoxic insults, ATM and related protein kinases phosphorylate the carboxyl-terminal tail of the H2AX protein (gamma-H2AX). gamma-H2AX marks the site of damage and provides a nucleation site for the formation of damage response and repair complexes. This modification, called gamma-H2AX (phospho-Ser139), triggers cell cycle arrest and DNA damage response repair, which makes this assay particularly appropriate for studying genome stability, cell cycle, DNA repair, and more broadly as a biomarker for cancer and for aging process investigations.

2019-08-22 · Over the past decade, the γ-H2AX assay has been applied to a variety of cell types and tissues to correlate γ-H2AX levels with DNA damage and repair [9,10,11,12,13]. Following radiation exposure, histone H2AX is rapidly phosphorylated by ATM and/or DNA-PK kinases at or near the vicinity of DNA DSB sites to form γ-H2AX [ 14 ]. HT gamma -H2AX Pharmacodynamic Assay Summary.
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2018-06-01 · Gamma-H2AX assay has proved useful for detecting DNA damage at doses of radiation down to 1 mGy and it is said to be 100 times more sensitive than other methods , . Reportedly the gamma-H2AX can be quantified either by immunoflourescence or flow cytometry [14] .

Cells are cultured in microplates, treated with agents that induce DNA damage or apoptosis, which stimulates H2A.X phosphorylation. munocytoﬂuorescence assay can be used to quantify ra-diation-induced changes in the phosphorylation status of the histone 2A.X (H2AX) (Redon et al. 2009). This ap-proach allows for radiation dose estimation from a mea-surement of cH2AX foci, as previously demonstrated in human cells and non-human primates (Redon et al.
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Supplemental Information. P-TEFb Activation by RBM7

Assuming that a single gamma-H2AX focus corresponds to a single DSB, one can convey the severity of cellular damage by the number of gamma-H2AX foci per cell (FPC) [1]. Thus, with γH2AX we have a bio-indicator on hand for critical DNA lesions and severe genomic replication stress. Hence, the γH2AX assay is not an assay for detecting any type of DNA damage, but for biologically relevant and severe DNA damage (including DNA interstrand crosslinks that give rise to DSBs; Nikolova et al., 2012). Phosphorylated H2AX (gamma-H2AX) is essential to the efficient recognition and (or) repair of DNA double strand breaks (DSBs), and many molecules, often thousands, of H2AX become rapidly phosphorylated at the site of each nascent DSB. An antibody to gamma-H2AX reveals that this highly amplified process generates nuclear foci. 2017-02-03 · 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.

Assaying DNA Damage in Hippocampal Neurons Using the

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2012 The focus of the study is an intercomparison of laboratories' dose-assessment performances using the y.