抗gamma H2AX [p Ser139]抗体 | Anti-gamma H2AX [p Ser139] Antibody

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	Immunocytochemistry/Immunofluorescence: gamma H2AX [p Ser139] Antibody [NB100-384] - Samples: Neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right). Antibody: Affinity purified rabbit anti-gamma-H2AX used at a dilution of 1:5,000 (0.2ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight 594 used at a dilution of 1:100.
	Flow Cytometry: gamma H2AX [p Ser139] Antibody [NB100-384] - Analysis of gamma-H2AX in Etoposide Treated Jurkat Cells. Cells were treated for 3 hrs in 5ug/ml etoposide, fixed in 1.5% PFA, and permeabilized in 90% Methanol. 1 million cells were stained with 0.5 ug anti-KLH or anti-H2AX NB100-384 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- etosposide treated, anti-KLH; Red- untreated, anti-gamma H2AX; Blue- etoposide treated, anti-gamma H2AX.
	Biological Strategies Validation. Western Blot: gamma H2AX [p Ser139] Antibody [NB100-384] - Samples: Nuclear extract from HeLa cells treated with 100 uM etoposide for 4 hours (+) or mock treated (-). Antibody: Affinity purified rabbit anti-gamma-H2AX antibody used at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
	Immunohistochemistry: gamma H2AX [p Ser139] Antibody [NB100-384] - Sample: FFPE section of human testicular seminoma. Antibody: Affinity purified rabbit anti- gamma-H2AX used at a dilution of 1:1,000 (1 ug/ml). Detection: DAB
	Immunohistochemistry: gamma H2AX [p Ser139] Antibody [NB100-384] - Samples: Neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right). Antibody: Affinity purified rabbit anti-gamma-H2AX used at a dilution of 1:5,000 (0.2ug/ml). Detection: Red fluorescent Anti-rabbit IgG-DyLight 594 conjugated used at a dilution of 1:100.
	Biological Strategies Validation. Western Blot: gamma H2AX [p Ser139] Antibody [NB100-384] - Detection of Human and Mouse gamma H2AX by Western Blot. Samples: Nuclear extract (50 ug) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: Affinity purified rabbit anti-gamma H2AX antibody NB100-384 used at 0.1 ug/ml. Detection: Chemiluminescence with 30 second exposure. (NCS, neocarzinostatin - 200 ng/ml, 30 min).
	Simple Western: gamma H2AX [p Ser139] Antibody [NB100-384] - Simple Western lane view shows a specific band for gamma H2AX in 0.2 mg/ml of Jurkat lysate(s). This experiment was performed under reducing conditions using the 12 - 230 kDa separation system.
	Simple Western: gamma H2AX [p Ser139] Antibody [NB100-384] - Electropherogram image(s) of corresponding Simple Western lane view. Gamma H2AX antibody was used at 5 ug/ml dilution on Jurkat lysate(s).

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Background

Histone H2AX is a 14 kDa ubiquitous member of the H2A histone family. It contains an evolutionary conserved SQ motif at the C-terminus in eukaryotes. Serine 139 within this motif becomes rapidly phosphorylated to yield a form known as gamma-H2AX in response to double-strand DNA damage. The response of eukaryotic cells to double-strand breaks in genomic DNA includes the sequestration of many factors into nuclear foci. H2AX becomes extensively phosphorylated within 1-3 minutes of DNA damage and forms foci at break sites.

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