Anti-gamma H2A.X (phospho S139) antibody [9F3]

• WB

Lab

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

Western blot : Mouse Monoclonal [9F3] to gamma H2A.X phospho S139 ab26350 staining at 1/1000 dilution, shown in green; Rabbit anti GAPDH (ab181602) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 15 kDa in HepG2 Vehicle Control Etoposide (0 uM, 90 min) cell lysates with no signal observed at this size in HepG2 Treated Etoposide (100 uM, 90 min) cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc BSA in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350) at 1/1000 dilution

Lane 1:

HepG2 Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 2:

HepG2 Treated Etoposide (100 uM, 90 min) at 20 µg

Lane 3:

Jurkat Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 4:

Jurkat Treated Etoposide (100 uM, 90 min) at 20 µg

Lane 5:

Human Brain at 20 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 15 kDa

Observed band size: 15 kDa

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• Flow Cyt

Unknown

Flow Cytometry - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

Overlay histogram showing HeLa cells stained with ab26350 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab26350, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG1 [ICIGG1], (ab91353, 1μg/1x10⁶ cells), IgG2a [ICIGG2A], (ab91361, 1μg/1x10⁶ cells), IgG2b [PLPV219], (ab91366, 1μg/1x10⁶ cells), IgG3 [MG3-35], (ab18394, 1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

• WB

Lab

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

Rabbit anti GAPDH was used as a GAPDH loading control. The intensity of the band has increased after treatment in both HepG2 and Jurkat cells.

All lanes:

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350) at 1/1000 dilution

Lane 1:

HepG2 Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 2:

HepG2 Treated Etoposide (100 uM, 90 min) at 20 µg

Lane 3:

Jurkat Vehicle Control Etoposide (0 uM, 90 min) at 20 µg

Lane 4:

Jurkat Treated Etoposide (100 uM, 90 min) at 20 µg

Lane 5:

Human Brain at 20 µg

Predicted band size: 15 kDa

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• WB

Unknown

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)

Lane 1:

Molecular weight marker

Lane 2:

Cell lysates prepared from Jurkat (Human T cell leukemia cell line from peripheral blood) cells

Lane 3:

Cell lysates prepared from Jurkat cells treated with staurosporine

Lane 4:

Cell lysates prepared from NIH/3T3 (Mouse embryonic fibroblast cell line) cells

Lane 5:

Cell lysates prepared from CHO-K1 (Chinese hamster ovary cell line) cells

Lane 6:

Cell lysates prepared from Rat2 (Rat fibroblast cell line) cells

Predicted band size: 15 kDa

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• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

H2A.X immunocytochemistry-immunofluorescence using Anti-gamma H2A.X (phospho S139) antibody [9F3] ab26350. Publication image and figure legend from Yan, S., Liu, L., et al., 2017, Cell Death Dis, 8 (8), Pubmed 28796254.

LC3 interacts with γ-H2AX and Rad51. (a and b) 786-O cells were treated with ST (0-8 μm) or rasfonin (0-6 μm) for 3 h, cells were lysed and subjected to immunoblotting with the antibodies indicated. (c) 786-O cells were treated with ST or rasfonin for 3 h, and the images were obtained using fluorescence microscopy following staining with the antibodies of LC3 and γ-H2AX. (d-f) 786-O cells were incubated with ST or rasfonin for 3 h and lysed, and LC3s or pULK1s were precipitated using the antibody against LC3 or pULK1 (Ser555). The immunoprecipitates were resolved by electrophoresis and probed by immunoblotting with the indicated antibodies. All data were acquired from three independent experiments

• WB

CiteAb

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

gamma H2A.X western Blotting using Anti-gamma H2A.X (phospho S139) antibody [9F3] ab26350. Publication image and figure legend from Yan, S., Liu, L., et al., 2017, Cell Death Dis, PubMed 28796254.

LC3 interacts with γ-H2AX and Rad51. (a and b) 786-O cells were treated with ST (0-8 μm) or rasfonin (0-6 μm) for 3 h, cells were lysed and subjected to immunoblotting with the antibodies indicated. (c) 786-O cells were treated with ST or rasfonin for 3 h, and the images were obtained using fluorescence microscopy following staining with the antibodies of LC3 and γ-H2AX. (d-f) 786-O cells were incubated with ST or rasfonin for 3 h and lysed, and LC3s or pULK1s were precipitated using the antibody against LC3 or pULK1 (Ser555). The immunoprecipitates were resolved by electrophoresis and probed by immunoblotting with the indicated antibodies. All data were acquired from three independent experiments

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• WB

CiteAb

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

-gamma H2A.X western blotting using Anti-gamma H2A.X (phospho S139) antibody [9F3] ab26350. Publication image and figure legend from Zhai, Q. Y., Ge, W., et al., 2018, Aging (Albany NY), PubMed 30153657.

nZnO induces DNA damage in fetal oocytes in vitro. (A) Representative double IF of pachytene and diplotene oocytes for SCP3 (red) and γH2AX (green) obtained from ovarian tissues cultured for 6 days. (B) Quantification of oocytes showing distinct γH2AX staining. (C) Representative WB for the indicated proteins from ovarian tissues cultured for 6 days; actin was used as loading control. (D) Representative double IF of pachytene and diplotene oocytes for SCP3 (red) and RAD51 (green). (E) Quantification of oocytes showing RAD51 foci.

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• WB

CiteAb

Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (AB26350)

gamma H2A.X western blotting using Anti-gamma H2A.X (phospho S139) antibody [9F3] ab26350. Publication image and figure legend from Li, J. L., Wang, J. P., et a., 2019, Cancer Med, PubMed 31670906.

FEN1 is overexpressed in cervical cancer and upregulated by ionizing radiation (IR) induction. A, Expression of FEN1 in cervical cancer samples (Tumor) vs control tissues (Ctrl), using the TCGA cervical cancer dataset. B, Scatter plots showing expression of FEN1 and γH2AX in cervical cancer samples. C, Cervical cancer samples were stratified based on median FEN1 expression. GSEA shows enrichment of the GNF2_H2AFX signature in FEN1High samples vs FEN1Low samples. D, FEN1 and γH2AX expression levels in HeLa cells were determined after 2 h of IR treatment. E FEN1 and γH2AX expression levels at different time point in HeLa cells were determined after IR (5 Gy) treatment

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