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Step : 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. Materials and methods: Whole blood and separated lymphocyte samples were homogenously irradiated with 60Co gamma rays at 0.5, 2.5 (blind samples), 0 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. There is a paucity of large animal models to study both the extent and the health risk of ionizing radiation exposure in humans.
19 Mar 2020 The γ-H2AX assay after external exposure is a good tool for However, less is known about DNA DSBs and γ-H2AX foci within the tissue 22 Aug 2019 Measurement of γ-H2AX foci levels in cells provides a sensitive and reliable method for quantitation of the radiation-induced DNA damage Phosphorylation of histone H2AX to form gamma-H2AX is a known marker for irradiation-induced DNA DSBs. However, the first generation assay involves the 28 Sep 2018 Thus, to apply the most suitable γH2AX analysis, specific assay characteristics rapid phosphorylation of the core histone variant H2AX at serine 139 is Use of the gamma-H2AX assay to monitor DNA damage and repair i 20 May 2016 Therefore, the combination of comet assay and gammaH2AX Characteristics of gamma-H2AX foci at DNA double-strand breaks sites. 20 Jan 2016 The questions that remain are whether the γ-H2AX assay can discriminate between cell strains with a known mutation in the DNA damage repair 30 Jun 2014 Several assays are routinely used to test genotoxicity in vitro, including the Ames assay, mouse lymphoma assay, micronucleus assay, and When the γ -H2AX foci which mark the DSBs are stained, individual breaks are detectible, making the assay suitable for situations requiring great sensitivity. Human gamma H2A.X (phospho S139) peptide X phospho S139 is used in this high-throughput duplexing plate-based assay. H2A.X (phospho S139) is a This modification, called gamma-H2AX (phospho-Ser139), triggers cell cycle arrest and DNA damage response repair, which makes this assay particularly the γ-H2AX Assay to Investigate DNA repair dynamics following multiple radiation exposures.
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. DNA double-strand break repair kinetics in minipig blood lymphocytes and fibroblasts, based on the γ-H2AX assay, were similar to those observed in their human counterparts.
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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.
A control for the day-to-day normalization of the flow cytometry
In vivo. 2008;22(3): 305-9. 10.
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
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.
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Se hela listan på academic.oup.com 2012-12-31 · The γ-H2AX assay has also been used to monitor formation and persistence of DNA damage and cytotoxicity in human cancer cells produced by another Auger emitter, 111 In, that has been delivered to cell nuclei by pharmaceuticals that target human epidermal growth factor receptor (EGFR)-positive cancers and HER2/neu-amplified breast cancers , . PDF | On Jan 1, 2008, I.M. Rakiman and others published γ-H2AX assay: a technique to quantify DNA double strand breaks | Find, read and cite all the research you need on ResearchGate Gamma-H2AX assay is an immuno-fluorescence experiment that enables detecting the location and number of DNA double strand breaks (DSBs) in cells. 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].
The instrument was used to image the stained DNA using the GFP imaging channel. Integrated Gen5™ Microplate Reader and Imager Software controls Cytation 5, and also automates image capture, analysis and processing. Methods. Gamma H2AX Assay Performance Cultured U251 cells were treated with
The DNA double-strand break (DSB) is the primary lethal lesion after therapeutic radiation.
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A control for the day-to-day normalization of the flow cytometry
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). CONCLUSIONS: This inter-comparison exercise has demonstrated that following a recent acute radiation exposure, the gamma-H2AX assay could be a useful triage tool that can be successfully applied across a network of laboratories. PMID: 27686523 [Indexed for MEDLINE] Publication Types: Comparative Study; Evaluation Studies; Validation Studies; MeSH terms 2013-07-08 · DNA double-strand break repair kinetics in minipig blood lymphocytes and fibroblasts, based on the γ-H2AX assay, were similar to those observed in their human counterparts. To substantiate the similarity observed between the human and minipig we show that minipig fibroblast radiosensitivity was similar to that observed with human fibroblasts. A number of ELISA Kits are available that target gammaH2ax, an alias of the protein, H2A histone family member X. The human protein, encoded by the gene H2AFX, is a member of the Histone H2A family. The protein's cellular localization is indicated to be nuclear.
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I stained pancreatic cancer cells (Panc-1) for gamma-H2AX and got a large amount of foci in control cells (cells which have not undergone any genotoxic stress 2013-07-08 CONCLUSIONS: This inter-comparison exercise has demonstrated that following a recent acute radiation exposure, the gamma-H2AX assay could be a useful triage tool that can be successfully applied across a network of laboratories. PMID: 27686523 [Indexed for MEDLINE] Publication Types: Comparative Study; Evaluation Studies; Validation Studies; MeSH terms gamma-H2AX Pharmacodynamic Assay Kit. The gamma-H2AX Pharmacodynamic Assay Kit is for the study of double strand DNA breaks through the detection of gamma-H2AX in peripheral blood mononuclear cells, cultured cells, and tissue biopsies. Contract Services are also available. 2013-11-29 Gamma-H2AX assay is an immuno-fluorescence experiment that enables detecting the location and number of DNA double strand breaks (DSBs) in cells. 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].
Evaluation of the gamma-H2AX assay for radiation biodosimetry in a swine model. Moroni M(1), Maeda D, Whitnall MH, Bonner WM, Redon CE. Author information: (1)Armed Forces Radiobiology Research Institute, Uniformed Services University, Bethesda, MD 20889-5603, USA. maria.moroni@usuhs.edu 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 proc … Although it is commonly accepted that a γ-H2AX focus indicates the presence of a double strand break (DSB), while foci disappearance is associated with the repair of the DNA damage, the exact relationship between the number of foci and the number of DSBs is still a matter of debate [ 12 - 14 ]. The γ-H2AX assay has been applied for a variety of human tissues and cells. The best way to monitor the efficiency of cancer therapy would be to directly assay tumor biopsies, although a high variability of responses can occur due to cellular heterogeneity and vascularization. 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 ].