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Detection of human gamma-H2AX by western blot. Samples: Whole cell lysate (15 µg) from Jurkat cells treated with 100 µM etoposide for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit anti-gamma-H2AX antibody A300-081A (lot A300-081A-19) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds. Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893) HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were incubated at 37°C for 3h with vehicle control (0 µM) and different concentrations of camptothecin ( ab120115 ). Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174) All lanes : Anti-gamma H2A.X (phospho S139) antibody (ab11174) at 0.04 µg/ml Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cells treated with 100 µM etoposide, whole cell lysate I have gotten good results with H2A.X and gH2A.X blots using antibodies from Cell Signaling. I don't do anything unusual for the western blot except use a 12% gel and be careful not to run H2A.X The attached is a γH2AX western blotting result.293T cells were treated with etoposide or DMSO (control),then cells were lysed in RIPA buffer with sonication.0.2 nitrocelluose membrane was used Gamma H2AX and its phosphorylation can be detected through western blotting and immunofluorescence, serving as a biomarker for DSB frequency (1-4).

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This experiment was performed under reducing conditions using the 12 - 230 kDa separation system. MA1-2022 detects human and mouse phosphorylated H2AX. MA1-2022 has been used successfully in Western blot, immunofluorescence, and ELISA procedures. By Western blot this antibody detects ~17 kDa protein representing phosphorylated H2AX in gamma irradiated HeLa cell lysate. Phospho gamma H2AX pSer139 Polyclonal Antibody for Western Blot IF ICC IHC P. Catalog Number: PA125001.

Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA is wrapped around histone-groups, consisting of the core histones H2A, H2B, H3 and H4. As a reaction on DNA Double-strand breaks (DSB) H2AX becomes phosphorylated on serine 139, called gamma-H2AX. MA1-2022 detects human and mouse phosphorylated H2AX.

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10856-1-AP. 2021-04-06 Phosphorylated H2AX (also termed, gamma-H2AX) functions to recruit and localize DNA repair proteins or cell cycle checkpoint factors to the DNA-damaged sites. In this way, phosphorylated H2AX promotes DNA repair and maintains genomic stability and thus helps prevent oncogenic transformations.

H2AX phosphorylation in A549 cells induced by the bulky and

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. The histone H2A.X is a variant member of the H2A family of histones that is distinguished from other H2A histones by a unique carboxy-terminal sequence. This unique sequence is highly conserved throughout eukaryotic evolution and is rapidly phosphorylated by ATM or ATR at Serine 139 in mammals in response to DNA double-strand breaks.

gamma H2A.X (phospho S139) antibody [3F2] Abcam. catalog: ab22551.
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We describe here an ELISA assay that Western Blot: gamma H2AX [p Ser139] Antibody [NB100-384] - RIPA lysate (50 and 15 mcg) from HeLa cells that were either treated with etoposide (60 minutes) or mock treated. Flow Cytometry: gamma H2AX Phosphospecific Antibody [NB100-384] - Flow Cytometrical Analysis of gamma-H2AX in Etoposide Treated Jurkat Cells.

Abcam H2ax antibody. select up to five products for comparison. gamma H2A.X (phospho S139) antibody [3F2] Abcam. catalog: ab22551.
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801 - 809 Article Download PDF View Record in Scopus Google Scholar 2010-03-06 · Background When DNA double-strand breaks (DSB) are induced by ionizing radiation (IR) in cells, histone H2AX is quickly phosphorylated into γ-H2AX (p-S139) around the DSB site. The necessity of DNA-PKcs in regulating the phosphorylation of H2AX in response to DNA damage and cell cycle progression was investigated. Results The level of γH2AX in HeLa cells increased rapidly with a peak level Anti-H2AX pS139 antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user.