I performed IF to find gamma H2AX foci in mammalian cell with it seems that I detect the exact same bands using total H2A.X antibody and phospho-H2AX (both antibodies from cell signaling

After the initial sensing of the DSB by MRN and the activation of ATM, H2AX is phosphorylated, which in turn elicits a sequence of signaling events thought to ultimately cause the activation of nucleases such as Mre11 and CtIP to process the DNA ends and generate ssDNA with 3′ overhangs. ssDNA is bound by RPA, which is subsequently exchanged by Rad51 and Rad51 paralogs.

γH2AX does not, itself, cause chromatin decondensation, but within 30 seconds of ionizing radiation , RNF8 protein can be detected in association with γH2AX. [13] H2A.X is required for checkpoint-mediated cell cycle arrest and DNA repair following double-stranded DNA breaks (1). DNA damage, caused by ionizing radiation, UV-light, or radiomimetic agents, results in rapid phosphorylation of H2A.X at Ser139 by PI3K-like kinases, including ATM, ATR, and DNA-PK (2,3). H2A.X is phosphorylated at Ser139 by DNA-PK in response to cell death receptor activation, c-Jun N-terminal Kinase (JNK1) in response to UV-A irradiation, and p38 MAPK in response to serum starvation (5-8).

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phosphorylated histone H2AX was predictive of disease progression epithelial dysplasia of the oral cavity. Rabbit Polyclonal Anti-gamma H2AX [p Ser139] Antibody DNA Double-strand break marker cited in 104 publications. Validated: WB, Simple Western, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, ChIP, KO. Within minutes of the induction of DNA double-strand breaks in somatic cells, histone H2AX becomes phosphorylated at serine 139 and forms gamma-H2AX foci at the sites of damage. These foci then play a role in recruiting DNA repair and damage-response factors and changing chromatin structure to accurately repair the damaged DNA. These gamma-H2AX foci appear in response to irradiation and Attenuation of genotoxicity under adhesion-restrictive conditions through modulation of p53, gamma H2AX and nuclear DNA organization.

to reduce the background continuum and to improve the signal-to-noise ratio.

Tekniker för att inducera och kvantifiera cellulärt åldrande som kännetecknas av utsöndring av ett antal pro-inflammatoriska cytokiner och signalmolekyler. Att kvantifiera antalet av γ-H2AX foci, räkna γ-H2AX foci per cell.

After the initial sensing of the DSB by MRN and the activation of ATM, H2AX is phosphorylated, which in turn elicits a sequence of signaling events thought to ultimately cause the activation of nucleases such as Mre11 and CtIP to process the DNA ends and generate ssDNA with 3′ overhangs. ssDNA is bound by RPA, which is subsequently exchanged by Rad51 and Rad51 paralogs. H2A.X is phosphorylated at Ser139 by DNA-PK in response to cell death receptor activation, c-Jun N-terminal Kinase (JNK1) in response to UV-A irradiation, and p38 MAPK in response to serum starvation (5-8). H2A.X is constitutively phosphorylated on Tyr142 in undamaged cells by WSTF (Williams-Beuren syndrome transcription factor) (9,10).

2003-02-01

The gamma-H2AX signal was quantitative with a linear increase of the gamma-H2AX signal over two orders of magnitude.

15% γ-H2AX is a specific and efficient coordinator of DDR signaling. Why does the cell initiate a spatially restricted modification of chromatin in the form of H2AX phosphorylation in response to DSBs? Which aspects of the DDR are facilitated through this modification? Wash sections in dH 2 O two times for 5 min each. Wash sections in wash buffer for 5 min. Block each section with 100–400 µl of preferred blocking solution for 1 hr at room temperature. Remove blocking solution and add 100–400 µl primary antibody diluted in SignalStain ® Antibody Diluent ( #8112) to each section.
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Strasberg Rieber M(1), Viola-Rhenals M, Rieber M. Author information: (1)IVIC, Tumor Cell Biology Laboratory, CMBC, Apartado 21827, Caracas, Venezuela.

Secondary antibody Mouse IgG-Fc (FITC), ab97264 , 1mg. 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). H2AX is a variant of histone H2A, one of the histone core molecules forming the nucleosome, and is a vital component in repairing DNA damage (1-4).
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Monoclonal Antibody for studying H2AX (Ser139) phosphate in the Chromatin Regulation / Epigenetics research area.

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). H2AX is a variant of histone H2A, one of the histone core molecules forming the nucleosome, and is a vital component in repairing DNA damage (1-4). Phosphorylated on Ser-140 (to form gamma-H2AX or H2AX139ph) in response to DNA double strand breaks (DSBs) generated by exogenous genotoxic agents and by stalled replication forks, and may also occur during meiotic recombination events and immunoglobulin class switching in lymphocytes. 2014-03-06 · HEK293T cells were transfected with H2AX and derivatives either (−) or (+) RNF168 and analyzed as in Figure 1D. (B) Human H2AX and derivatives reconstituted in MCF10A H2AX −/− cells.