an X93-FITC filter was used to detect anti-γ-H2AX and anti-phospho-histone H3 antibodies, and an X93-Cy5 filter was used to detect anti-tubulin antibody. A 10× objective lens was used to assess the cell cycle (Hoechst 33342), and a 20× high NA objective lens was used to visualize anti-γ-H2AX, anti-phospho-histone H3, and anti-tubulin staining.

Co-staining-proceduren för γ-H2AX och 53BP1 är ett standardförfarande för detektion av DNA-DSB. Bildandet av γ-H2AX foci är associerad 

by. Blue staining indicates cell nuclei while pink staining indicates cytoplasm. (Other panels) γ-H2AX staining in tissues irradiated with 0, 0.2, 2.0, and 5.0 Gy and fixed   By coupling γ-H2AX immunofluorescent labeling with either PI (flow cytometry) or DAPI (DIM) staining of the DNA, we were able to easily discern distinct cell cycle   A, γ irradiation dosage dependent staining of γ-H2AX on T cells. DNA breaks were generated by γ irradiation with a Cs137 source, and cells were stained 20 min  γ-H2AX staining was not attributable to host DNA damage, as the host genome was found to be intact. The response was rather a result of overactivated ATR,  16 Aug 2012 4.1 Immunofluorescence staining of γH2AX in vitro. phosphorylated H2AX ( γH2AX), which form foci covering many megabases of chromatin  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  The scoring of p-H2AX staining was based on intensity grade representing the In the later stages of DDR, the γ-H2AX foci become larger and, thus, a higher  28 Sep 2018 Quantification of H2AX levels by immunoblotting was based on the protocol Samples with >5% of cells showing a pan-nuclear γH2AX staining were Use of the gamma-H2AX assay to monitor DNA damage and repair in  Gamma H2AX (gammaH2AX) is the phosphorylated version of histone H2AX and is a marker for double-stranded breaks (DSBs) caused by DNA damage (1-4). 2.4.

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Moreover, since gamma‐H2AX staining can be performed on formalin‐fixed and paraffin‐embedded tissue sections generated during repeated‐dose toxicity studies, it does not require any further treatments or extra procedures during dissection, thus optimizing the use of resources and animals. A fragment of each xenograft was fixed and processed for γ-H2AX immunofluorescent staining (IFA). γ-H2AX staining is green; nuclei are stained with DAPI blue. (C-E) Bar graphs showing quantification of γ-H2AX IFA staining (C), % γ-H2AX ELISA readings (D), and PAR ELISA readings (E) in xenograft tumors. Mice were treated as indicated on the bottom of each figure. Abstract UV irradiation induces histone variant H2AX phosphorylated on serine 139 (γH2AX) foci and high levels of pan-nuclear γH2AX staining without foci, but the significance of this finding is still uncertain. We examined the formation of γH2AX and 53BP1 that coincide at sites of double-strand breaks (DSBs) after ionizing radiation.

γ-H2AX staining is green; nuclei are stained with DAPI blue. (C-E) Bar graphs showing quantification of γ-H2AX IFA staining (C), % γ-H2AX ELISA readings (D), and PAR ELISA readings (E) in xenograft tumors.

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, 

BioAssay record AID 1254631 submitted by ChEMBL: Induction of DNA damage in human HCT116 cells at G0/G1 phase at 30 uM after 24 hrs by gamma-H2AX-staining based-flow cytometry. 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 Phosphorylation of the chromatin protein H2AX (forming γH2AX) is implicated in the repair of DNA double strand breaks (DSB's); a large number of H2AX molecules become phosphorylated at the sites of DSB's. Fluorescent staining of the cell nuclei for γH2AX, via an antibody, visualises the formation of these foci, allowing the quantification of DNA DSB's and forming the basis for a sensitive Rabbit Polyclonal Anti-gamma H2AX [p Ser139] Antibody DNA Double-strand break marker cited in 104 publications.

The results suggest gamma-H2AX is a useful adjunct in diagnosis of metastatic RCC when RCC-Ma is negative and in higher grade RCC, which are often a diagnostic challenge. A nuclear pattern of staining of gamma-H2AX has a comparable sensitivity with RCC-Ma, and the interpretation is easier and more reliable. RCC-Ma is 100% specific for RCC, but only when a membranous pattern of staining is interpreted as positive. PMID: 18528282 [Indexed for MEDLINE]

Pan-nuclear staining of γ-H2AX is also known to have different size and intensity based on the type of radiation exposure 14,23.Particle based irradiation response such as that induced by heavy Positive immunohistochemical staining of gammaH2AX is associated with tumor progression in gastric cancers from radiation-exposed patients. Sentani K (1), Oue N, Sakamoto N, Nishisaka T, Fukuhara T, Matsuura H, Yasui W. Author information: (1)Department of Molecular Pathology, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan. These observations suggest that the two methods could complement each other for DNA damage analysis, where gamma‐H2AX staining allows the detection of tissue‐specific effects in situ. Moreover, since gamma‐H2AX staining can be performed on formalin‐fixed and paraffin‐embedded tissue sections generated during repeated‐dose toxicity studies, it does not require any further treatments or extra procedures during dissection, thus optimizing the use of resources and animals. A fragment of each xenograft was fixed and processed for γ-H2AX immunofluorescent staining (IFA). γ-H2AX staining is green; nuclei are stained with DAPI blue.

Moreover, since gamma‐H2AX staining can be performed on formalin‐fixed and paraffin‐embedded tissue sections generated during repeated‐dose toxicity studies, it does not require any further treatments or extra procedures during dissection, thus optimizing the use of resources and animals. 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.
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Fluorescent staining of the cell nuclei for γH2AX, via an antibody, enables us to visualise the formation of these foci, allowing the quantification of DNA DSB’s This expression was defined as γ-H2AX domains at leptotene and zygotene stages whose intensity declined on autosomes in relation with synapsis formation. Then, from late zygotene to diplotene γ-H2AX staining exclusively involved the sex body. Moreover, since gamma‐H2AX staining can be performed on formalin‐fixed and paraffin‐embedded tissue sections generated during repeated‐dose toxicity studies, it does not require any further treatments or extra procedures during dissection, thus optimizing the use of resources and animals. Histone H2AX Antibody Staining Protocol for Immunohistochemistry . Description: Histone H2AX is a 14 kDa ubiquitous member of the H2A histone family that contains an evolutionary conserved SQ motif at the C-terminus in eukaryotes.

spermatogonia with γ-H2AX immunoflourescence staining after radionuclide uptake in the mouse testis – method development with Indium-111  Developmental effects of fractionated low-dose exposure to gamma radiation on Long time persistence of residual 53BP1/gamma-H2AX foci in human  (författare); Quantitative γ-H2AX immunofluorescence method for DNA double-strand break analysis in testis and liver after intravenous administration of  immediately after the last administration. Immunofluorescence staining for phosphorylation of histone H2AX assay on bone marrow cells. Eftersom överflöd av γ H2AX indikerar omfattningen av DSB, identifierade vi först ( d ) Immunostaining-analys av olika faser av cellcykel av MRC-5-celler.
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(C) (Left) Representative DAPI and anti-g-h2ax staining images of HeLa cells 95 GAMMA: 3.45k, 59.5k, 118.8k/TM119 Condition D GAMMA SOURCE: Co6 

γ-H2AX staining is green; nuclei are stained with DAPI blue. (C-E) Bar graphs showing quantification of γ-H2AX IFA staining (C) , % γ-H2AX ELISA readings (D) , and PAR ELISA readings (E) in xenograft tumors. 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).

TSB treatment and staining; RNA fluorescence in situ hybridization (FISH) on av Ki67 eller fosforylerad gamma H2AX histon (yH2AX) i MAoEC behandlade 

an X93-FITC filter was used to detect anti-γ-H2AX and anti-phospho-histone H3 antibodies, and an X93-Cy5 filter was used to detect anti-tubulin antibody. A 10× objective lens was used to assess the cell cycle (Hoechst 33342), and a 20× high NA objective lens was used to visualize anti-γ-H2AX, anti-phospho-histone H3, and anti-tubulin staining.

Nuclear staining by gamma-H2AX had a similar sensitivity of 70% for RCC but a lower specificity of 77%, as it was seen in 1 of 18 HCC (5%) and 8 of 21 (38%)ACC. In metastatic RCC, 83% (39/47) of tumors with a higher nuclear grade stained with gamma-H2AX, compared with 46% (11/24) of low nuclear grade (equivalent of Fuhrman 2 and lower) tumors. Flow cytometry measurement for gamma-H2AX.