Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)](https://www.abcam.com/ps/products/26/ab26350/Images/ab26350-278306-anti-gamma-h2ax-phospho-s139-antibody-9f3-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)")
Key features and details
- Mouse monoclonal [9F3] to gamma H2A.X (phospho S139)
- Suitable for: Flow Cyt, WB, IHC-P
- Reacts with: Mouse, Rat, Human, Chinese hamster
- Isotype: IgG
Overview
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Product name
Anti-gamma H2A.X (phospho S139) antibody [9F3]
See all gamma H2A.X primary antibodies -
Description
Mouse monoclonal [9F3] to gamma H2A.X (phospho S139) -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Chinese hamster -
Immunogen
Synthetic peptide corresponding to Human gamma H2A.X (phospho S139).
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Positive control
- WB: Staurosporine treated Jurkat cell lysate. NIH/3T3, CHO-K1 and Rat2 cell lysate. IHC-P: Human placenta and spleen tissue. Flow Cyt: HeLa cells.
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General notes
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. -
Storage buffer
Preservative: 0.09% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Purified from TCS. -
Clonality
Monoclonal -
Clone number
9F3 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab26350 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt |
Use at an assay dependent concentration.
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| WB | (6) |
Use at an assay dependent concentration. Detects a band of approximately 16 kDa.
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| IHC-P | (12) |
Use at an assay dependent concentration.
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| Notes |
|---|
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Flow Cyt
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Detects a band of approximately 16 kDa. |
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IHC-P
Use at an assay dependent concentration. |
Target
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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. -
Sequence similarities
Belongs to the histone H2A family. -
Developmental stage
Synthesized in G1 as well as in S-phase. -
Domain
The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family. -
Post-translational
modificationsPhosphorylated on Ser-140 (to form gamma-H2AFX 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. Phosphorylation can extend up to several thousand nucleosomes from the actual site of the DSB and may mark the surrounding chromatin for recruitment of proteins required for DNA damage signaling and repair. Widespread phosphorylation may also serve to amplify the damage signal or aid repair of persistent lesions. Phosphorylation of Ser-140 (H2AX139ph) in response to ionizing radiation is mediated by both ATM and PRKDC while defects in DNA replication induce Ser-140 phosphorylation (H2AX139ph) subsequent to activation of ATR and PRKDC. Dephosphorylation of Ser-140 by PP2A is required for DNA DSB repair. In meiosis, Ser-140 phosphorylation (H2AX139ph) may occur at synaptonemal complexes during leptotene as an ATM-dependent response to the formation of programmed DSBs by SPO11. Ser-140 phosphorylation (H2AX139ph) may subsequently occurs at unsynapsed regions of both autosomes and the XY bivalent during zygotene, downstream of ATR and BRCA1 activation. Ser-140 phosphorylation (H2AX139ph) may also be required for transcriptional repression of unsynapsed chromatin and meiotic sex chromosome inactivation (MSCI), whereby the X and Y chromosomes condense in pachytene to form the heterochromatic XY-body. During immunoglobulin class switch recombination in lymphocytes, Ser-140 phosphorylation (H2AX139ph) may occur at sites of DNA-recombination subsequent to activation of the activation-induced cytidine deaminase AICDA. Phosphorylation at Tyr-143 (H2AXY142ph) by BAZ1B/WSTF determines the relative recruitment of either DNA repair or pro-apoptotic factors. Phosphorylation at Tyr-143 (H2AXY142ph) favors the recruitment of APBB1/FE65 and pro-apoptosis factors such as MAPK8/JNK1, triggering apoptosis. In contrast, dephosphorylation of Tyr-143 by EYA proteins (EYA1, EYA2, EYA3 or EYA4) favors the recruitment of MDC1-containing DNA repair complexes to the tail of phosphorylated Ser-140 (H2AX139ph).
Monoubiquitination of Lys-120 (H2AXK119ub) by RING1 and RNF2/RING2 complex gives a specific tag for epigenetic transcriptional repression. Following DNA double-strand breaks (DSBs), it is ubiquitinated through 'Lys-63' linkage of ubiquitin moieties by the E2 ligase UBE2N and the E3 ligases RNF8 and RNF168, leading to the recruitment of repair proteins to sites of DNA damage. Monoubiquitination and ionizing radiation-induced 'Lys-63'-linked ubiquitination are distinct events. -
Cellular localization
Nucleus. Chromosome. - Information by UniProt
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Database links
- Entrez Gene: 3014 Human
- Entrez Gene: 15270 Mouse
- Entrez Gene: 500987 Rat
- Omim: 601772 Human
- SwissProt: P16104 Human
- SwissProt: P27661 Mouse
- Unigene: 477879 Human
- Unigene: 245931 Mouse
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Alternative names
- H2A histone family member X antibody
- H2A histone family member X antibody
- H2A.FX antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)This image is courtesy of an anonymous Abreview.
Ab26350 staining H2A.X in Human placenta tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% BSA for 30 minutes at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in TBS) for 16 hours at 4°C. A diluted Biotin conjugated Goat anti-mouse polyclonal (1/200) was used as the secondary antibody.
This image was generated using the ascites version of the product.
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![Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)](https://www.abcam.com/ps/products/26/ab26350/Images/ab26350-37297-anti-gamma-h2ax-phospho-s139-antibody-9f3-western-blot.jpg)
Western blot - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)All lanes : 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) cellsThis image was generated using the ascites version of the product.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)](https://www.abcam.com/ps/products/26/ab26350/Images/ab26350-37299-anti-gamma-h2ax-phospho-s139-antibody-9f3-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)ab26350 (1µg/ml) staining gamma H2A in human spleen, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining .
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.This image was generated using the ascites version of the product.
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![Flow Cytometry - Anti-gamma H2A.X (phospho S139) antibody [9F3] (ab26350)](https://www.abcam.com/ps/products/26/ab26350/Images/ab26350-37301-anti-gamma-h2ax-phospho-s139-antibody-9f3-flow-cytometry.jpg)
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (294)
ab26350 has been referenced in 294 publications.
- Jenster LM et al. P38 kinases mediate NLRP1 inflammasome activation after ribotoxic stress response and virus infection. J Exp Med 220:N/A (2023). PubMed: 36315050
- Muciño-Hernández G et al. Nucleophagy contributes to genome stability through degradation of type II topoisomerases A and B and nucleolar components. J Cell Sci 136:N/A (2023). PubMed: 36633090
- Zhu M et al. KYA1797K, a Novel Small Molecule Destabilizing β-Catenin, Is Superior to ICG-001 in Protecting against Kidney Aging. Kidney Dis (Basel) 8:408-423 (2022). PubMed: 36466073
- Wang W et al. Studying the mechanism of sperm DNA damage caused by folate deficiency. J Cell Mol Med 26:776-788 (2022). PubMed: 34953021
- Wu H et al. A homozygous loss-of-function mutation in FBXO43 causes human non-obstructive azoospermia. Clin Genet 101:55-64 (2022). PubMed: 34595750