Name: Anti-gamma H2A.X (phospho S139) antibody
SKU: ab2893
Rating: 4 (36 reviews)

Anti-H2A.X (S139 phospho) antibody is a rabbit polyclonal antibody that is used in gamma H2A.X western blot (WB), and immunocytochemistry/immunofluorescence (ICC/IF). Suitable for human, mouse and rat samples.

- Cited in over 390 publications
- Tried and trusted by researchers since 2004
- Affinity purified

Images

Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (AB2893), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Form: Liquid
Clonality: Polyclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	ICC/IF	WB
Human	Tested	Tested
Mouse	Expected	Tested
Rat	Expected	Tested
Chimpanzee	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 0.1 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1 µg/mL	Notes -
Species Rat	Dilution info 1 µg/mL	Notes -
Species Human	Dilution info 1/1000	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Chimpanzee	Dilution info -	Notes -

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Target data

See full target information H2AX phospho S139 + S139

Function

Variant histone H2A which replaces conventional H2A in a subset of nucleosomes. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. Required for checkpoint-mediated arrest of cell cycle progression in response to low doses of ionizing radiation and for efficient repair of DNA double strand breaks (DSBs) specifically when modified by C-terminal phosphorylation.

Alternative names

H2AFX, H2AX, Histone H2AX, H2a/x, Histone H2A.X

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Form: Liquid
Clonality: Polyclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Purification technique: Affinity purification Immunogen
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-gamma H2A.X (phospho S139) antibody (ab2893) is a rabbit polyclonal antibody and is validated for use in ICC/IF, WB in human, mouse, rat samples.

Anti-gamma H2A.X (phospho S139) antibody (ab2893) has been cited over 394 times in peer reviewed journals and is trusted by the scientific community.

Abcam's high quality validation processes ensure Anti-gamma H2A.X (phospho S139) antibody (ab2893) has high sensitivity and specificity.

Anti-gamma H2A.X (phospho S139) antibody (ab2893) has 32 independent reviews from customers.

Anti-gamma H2A.X (phospho S139) antibody (ab2893) specifically detects gamma H2A.X Phospho-S139 (UniProt ID: P16104; Molecular weight: 15kDa) and is sold in 50 µg selling sizes.

ab2893 is batch tested in peptide array, western blot and ICC only, although some customers have successfully used this product in IHC and ChIP (see images below). We would recommend Anti-gamma H2A.X (phospho S139) antibody [EP854(2)Y] ab81299 as an alternative product for use in IHC and ChIP.

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

If you do not find a host species to meet your needs, our catalogue and custom Chimeric range provides scientists the specificity of Abcam's RabMAbs in the species backbone of your choice. Remember to also review our range of edited cell lines, proteins and biochemicals relevant to your target that may help you further your research goals.

Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Gamma H2A.X also known as phospho H2A.X or ?H2A.X is a phosphorylated form of the histone variant H2A.X. It has a molecular weight of about 14 kilodaltons and occurs primarily in they nucleus. When DNA double-strand breaks (DSBs) occur serine 139 in H2A.X undergoes rapid phosphorylation resulting in gamma H2A.X. This modification happens swiftly at the site of damage and gamma H2A.X spreads over a large chromatin area facilitating the recruitment of DNA repair proteins. Gamma H2A.X staining typically evaluated using gamma H2A.X immunofluorescence techniques aids in identifying the presence and extent of DNA damage.

Biological function summary

Gamma H2A.X plays a role in DNA damage response and repair. It does not operate alone; it acts as part of a complex with other repair proteins. The formation of gamma H2A.X foci at DNA damage sites creates a signal attracting repair factors that help maintain genome stability. Its interaction with MDC1 and ATM proteins exemplifies its significant role in orchestrating an effective response to DNA damage. Beyond DNA repair gamma H2A.X influences cell cycle checkpoints permitting cells to pause and repair before proceeding with division.

Pathways

Gamma H2A.X plays a pivotal role in the DNA damage response (DDR) pathway. This pathway is essential for detecting and repairing DNA lesions to uphold genomic integrity. Within the DDR pathway gamma H2A.X is closely associated with proteins such as NBS1 and BRCA1 which assist in repairing double-strand breaks. In addition gamma H2A.X is integral to the ATM-ATR signaling pathway where its activation promotes cell survival following genotoxic stress by signaling for damage repair or triggering apoptosis.

Associated diseases and disorders

Gamma H2A.X has connections to cancer and neurodegeneration. Aberrant DNA repair pathways often indicated by persistent gamma H2A.X signals correlate with tumor formation and progression. For instance a failure to repair DNA damage effectively can lead to mutations that drive cancer development. Gamma H2A.X also links to neurodegenerative diseases where dysregulated DNA repair contributes to neuronal cell death. Proteins like p53 which regulate cell cycle and apoptosis further connect to gamma H2A.X bridging its role in disease pathogenesis.

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14 product images

Garcia, C.P. et al PLoS One. 2016; 11(3): e0152607. Fig.2a Published online 2016 Mar 31. doi: 10.1371/journal.pone.0152607 Reproduced under the Creative Commons licence https://creativecommons.org/licenses/by/4.0/
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
Immunofluorescence photomicrographs of genotoxic-treated (1μM during 3 h) human induced pluripotent stem cells (hiPSCs) and hESCs-derived neuroprogenitors (NP) performed immediately after CPT treatment (1μM during 3 h). The figure shows representative images of cells stained with primary antibodies against ATM phospho-serine1981 (pATM), histoneγH2AX, p53, p53 phospho serine 15 (p53pSer15). Nuclei were counterstained with DAPI. The scale bars represent 100 μm.
Histone gamma H2A.X was detected using ab2893.
From Figure 2a of Garcia et al PLoS One. 2016; 11(3): e0152607. Published online 2016 Mar 31. doi: 10.1371/journal.pone.0152607' PMID: 27030982
Reproduced under the Creative Commons licence: https://creativecommons.org/licenses/by/4.0/
Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
Gel type: MES
Blocking buffer: 2% BSA block
All lanes: Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1 µg/mL
Lane 1: NIH 3T3 nuclear lysate (triton enriched) at 10 µg
Lane 2: PC12 nuclear lysate (triton enriched) at 10 µg
Secondary
All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa
Exposure time: 4min
Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
All lanes: Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1/500 dilution
Lanes 1 and 3: Control HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate Histone preparation
Lanes 2 and 4: Colcemid treated HeLa whole cell lysate Histone preparation
Lane 5: Control HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate Histone preparation with Human Histone H2A.X (unmodified ) peptide (ab15646)
Lane 6: Colcemid treated HeLa whole cell lysate Histone preparation with Human Histone H2A.X (unmodified ) peptide (ab15646)
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa, 50 kDa
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining gamma H2A.X (phospho S139) in HeLa UV cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab2893 at 0.1µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
This image is courtesy of Kirk McManus
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
Asynchronous HeLa cells were paraformaldehyde fixed and immunofluorescently labeled with ab2893 that had been preincubated with either 1) non-phosphorylated or 2) phosphorylated H2AX peptide. Identical exposure times were employed. The Merge images present the DAPI and ab2893 channels as red and green, respectively. Scale bars represent 5μm.
1) Non-phosphorylated peptides
2) Phosphorylated peptides
Image courtesy of Dr. Kirk McManus
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
Asynchronous HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were exposed to 2Gy and permitted to recover for 30min. Cells were paraformaldehyde fixed (4%), immunofluorescently labeled with ab2893 and counterstained with DAPI. The merge image presents the DAPI and ab2893 channels as red and green, respectively. The scale bar represents 5μm.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2A.X in weri cells treated with TMPyP4 tosylate (TMPyP4 tosylate, Telomerase inhibitor ab120793), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of TMPyP4 tosylate, as described in literature.
The cells were incubated at 37°C for 24 hours in media containing different concentrations of TMPyP4 tosylate, Telomerase inhibitor ab120793 (TMPyP4 tosylate ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2A.X in MALME-3M cells treated with terfenadine (Terfenadine, K+ channel blocker. H1 antagonist. ab120270), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of terfenadine, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of Terfenadine, K+ channel blocker. H1 antagonist. ab120270 (terfenadine) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with CPT 11 (Irinotecan) (CPT 11 (Irinotecan), DNA topoisomerase I inhibitor ab141107), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of CPT 11 (Irinotecan), as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of CPT 11 (Irinotecan), DNA topoisomerase I inhibitor ab141107 (CPT 11 (Irinotecan)) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with SN 38 (SN 38, DNA topoisomerase I inhibitor ab141108), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of SN 38, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of SN 38, DNA topoisomerase I inhibitor ab141108 (SN 38) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 10-Hydroxycamptothecin (10-Hydroxycamptothecin, DNA topoisomerase I inhibitor ab141071), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of 10-Hydroxycamptothecin, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of 10-Hydroxycamptothecin, DNA topoisomerase I inhibitor ab141071 (10-Hydroxycamptothecin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
ab2893 staining γH2AX (phospho S139) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with camptothecin (Camptothecin, DNA topoisomerase inhibitor ab120115), by ICC/IF. Increased nuclear expression of γH2AX (phospho S139) correlates with increased concentration of camptothecin, as described in literature.
The cells were incubated at 37°C for 3h in media containing different concentrations of Camptothecin, DNA topoisomerase inhibitor ab120115 (camptothecin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
Functional Studies - 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 (Camptothecin, DNA topoisomerase inhibitor ab120115). Increased expression of γH2A.X (phospho S139) in HeLa cells correlates with an increase in camptothecin concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab2893 at 1 μg/ml and Anti-Histone H2A.X antibody - Nuclear Marker ab10475 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution and visualised using ECL development solution.
Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893)
Gel type : MES
Blocking buffer : 2% BSA block
All lanes: Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1 µg/mL
Lane 1: NIH/3T3 (mouse embryonic fibroblast cell line) nuclear lysate (triton enriched) at 10 µg
Lane 2: PC-12 (rat adrenal gland pheochromocytoma cell) nuclear lysate (triton enriched) at 10 µg
Secondary
All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 15 kDa
Observed band size: 17 kDa
Exposure time: 16min