Phosphorylation of histone protein H2AX on serine 139 (gamma-H2AX) occurs at sites flanking DNA double-stranded breaks (DSBs) and can provide a measure of the number of DSBs within a cell. We

Cells were cytospunned and stored at -20°C until immunostaining. 2.3.2 γ-H2AX immunostaining and analysis. Three slides by dose by experiment were stained 

Validated: WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, ChIP, KO. mosomal ends that are detectible by means of gamma-H2AX staining procedures (Li et al. 2011). Mammalian cells employ vigorous repair processes, including non-homologousend-joining(NHEJ),toreconnectbrokenchro-mosome ends, usually correctly, within a day or two after irradiation. Still the rejoining process is not perfect as has H2AX phosphorylation at Ser139 (formation of γ-H2AX) is an indicator of double-strand breaks in DNA (DSBs) after the action of different genotoxic stresses, including ionizing radiation, environmental agents, and chemotherapy drugs.

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15 May 2019 further measurement of the γ-H2AX fluorescence intensity and foci formation ( Figure 2D). Pan-. 8 nuclear γ-H2AX stained cells displayed a  15 May 2019 Images displayed here show cells in bright field, γ-H2AX staining, γ-H2AX foci mask, DRAQ5 nuclear staining and a merged image panel of  Cells were cytospunned and stored at -20°C until immunostaining. 2.3.2 γ-H2AX immunostaining and analysis. Three slides by dose by experiment were stained  9 Jun 2020 immunofluorescence procedure of γ-H2AX staining. However, it includes all details regarding the selection of appropriate antibodies with  22 Aug 2019 For the compensation, irradiated blood cells were stained with γ-H2AX antibody or DRAQ5 only and captured using the 488 nm laser without  Phosphorylation of the histone protein H2AX to form γ-H2AX foci directly represents 0-100 μM); γ-H2AX (green) and nuclear DNA stained with DAPI ( blue).

A breast cancer specimen that expresses endogenous γH2AX was also utilized as  14 Sep 2010 carcinoma-bearing mice, the values of total γ-H2AX staining were threefold higher at the base of the crypts, where proliferating cells are located  21 Jun 2013 quantifying γ-H2AX nuclear fluorescence for uniformly irradiated cell DNA double strand breaks visualized via γ-H2AX staining in cultured  27 Apr 2009 described a novel γ-H2AX nuclear staining, which we referred to as “the apoptotic γ-H2AX ring”. In this extra-view, we will focus on this novel  30 Oct 2006 γH2AX phosphorylated histone H2AX.

Phosphorylation of the Ser-139 residue of the histone variant H2AX, forming γH2AX, is an early cellular response to the induction of DNA double-strand breaks. Detection of this phosphorylation

by. Blue staining indicates cell nuclei while pink staining indicates cytoplasm.

tissues. gamma-H2AX has already been investigated in a variety of cancer types, including breast, lung, colon, cervix, and ovary cancers. The prognostic value of gamma-H2AX is indicated in certain cancer types, such as breast or endometrial cancer, but further investigation is needed to establish gamma-H2AX as a prognostic marker.

One possible application of the present method could also be in radiation-protection for in-vivo dosimetry after accidental exposure to radiation.

In parallel, staining for gammaH2AX was performed to visualise the residual foci. RESULTS: In the CFA, FaDu showed a higher radioresistance than SKX. After analysis of the residual foci data, we constructed 'predicted' survival curves using two different methods.
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CAS Article 2006-11-13 · Additionally, γH2AX (the phosphorylated form of H2AX) staining in spermatozoa persisted despite the fact of a decrease in the number of γH2AX foci in FL cells after H 2 O 2 removal. Collectively, these results demonstrate that oxidative stress can induce H2AX phosphorylation in human spermatozoa through DSB induction, and that γH2AX may be used as a sensitive, novel marker for such DSBs.

In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Gamma H2A.X Staining Kit (ab242296) is based on the phosphorylation of the histone H2A.X at serine 139 in response to DNA damaging agents which cause double strand breaks in cells that are cultured in microtiter plates. The kit provides sufficient reagents for up to 100 stainings in 96- well plate.
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In situ detection of DNA double strand breaks by immunofluorescent gamma-H2AX staining in mice exposed to multiwalled carbon nanotubes. Aimonen, K.

The sites of DSBs can be visualized as focal sites of γ-H2AX using antibodies and immunofluorescence microscopy. Listed are ELISA Kits for the detection of H2AX, an alias name of H2A histone family member X. The human protein, encoded by the gene H2AFX, is 143 amino acid residues long and has a mass of 15,145 daltons. Immunofluorescent staining and bioimaging analysis of cultured cells can be used to readily identify H2AX (pS139)-containing foci. As such, H2AX (pS139) immunofluorescence localization serves as a biomarker for nuclear sites of DNA damage (e.g., double-stranded DNA breaks) in affected cells. Immunofluorescent staining of human cell line. 15 May 2019 further measurement of the γ-H2AX fluorescence intensity and foci formation ( Figure 2D). Pan-.

An immunofluorescent staining method was employed to determine the levels of γ-H2AX (phosphorylated H2AX, a variant of the H2A protein family, which is a 

H2A.X Phosphorylation Assay Kit (Flow Cytometry) The H2A.X Phosphorylation Assay Kit (Flow cytometry) is a cell based assay formatted for flow cytometric detection of levels of phosphorylated Histone H2A.X.; find Sigma-Aldrich-17-344 MSDS, related peer-reviewed papers, technical documents, similar products & more at Sigma-Aldrich.

γ-H2AX foci are considered as early DNA DSB signalling both on hamster and human PCC (Figure 1A). On the same PCC cells, the FISH staining of telomere and  22 Feb 2017 were used as positive controls for gamma-H2AX staining. A breast cancer specimen that expresses endogenous γH2AX was also utilized as  14 Sep 2010 carcinoma-bearing mice, the values of total γ-H2AX staining were threefold higher at the base of the crypts, where proliferating cells are located  21 Jun 2013 quantifying γ-H2AX nuclear fluorescence for uniformly irradiated cell DNA double strand breaks visualized via γ-H2AX staining in cultured  27 Apr 2009 described a novel γ-H2AX nuclear staining, which we referred to as “the apoptotic γ-H2AX ring”. In this extra-view, we will focus on this novel  30 Oct 2006 γH2AX phosphorylated histone H2AX. DSB of γ-H2AX in cancer cell lines. immunofluorescent staining with a custom-made rabbit γH2AX. 1 Mar 2006 For BrdU and γ-H2AX double staining, cells were first incubated with 100 μg/ml of BrdU (Sigma) for 20 min, then 15 min after irradiation, were  25 May 2011 Here, we evaluated γ-H2AX formation in the adult rat brain following the nucleus with γ-H2AX staining) were also excluded from the analysis,  9 Dec 2011 Nuclei were stained with DAPI (blue), γ-H2AX foci are colored in In plant cells, formation of γ-H2AX foci at irradiation-induced DSBs has been  14 Apr 2015 Immunofluorescence staining of markers of a DNA damage response: 53BP1 ( green) and gamma-H2AX (red) in human osteoblast-like U-2 OS  and assessed for 7-H2AX focus formation by immunofluores- cence analysis.