Anti-gamma H2A.X (phospho S139) antibody (ab11174)
Key features and details
- Rabbit polyclonal to gamma H2A.X (phospho S139)
- Suitable for: IHC-P, ICC/IF, WB, Flow Cyt
- Reacts with: Mouse, Human
- Isotype: IgG
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Overview
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Product name
Anti-gamma H2A.X (phospho S139) antibody
See all gamma H2A.X primary antibodies -
Description
Rabbit polyclonal to gamma H2A.X (phospho S139) -
Host species
Rabbit -
Specificity
Using IF, this antibody was shown to bind to a non-nuclear location in Hela cells. -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF MouseHumanIHC-P HumanWB Human -
Immunogen
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General notes
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
Antibodies were affinity purified using the peptide immobilized on solid support. -
Clonality
Polyclonal -
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 ab11174 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
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ICC/IF |
Mouse
Human
|
IHC-P |
Human
|
WB |
Human
|
All applications |
Rabbit
Guinea pig
Cow
Dog
Pig
Rhesus monkey
Gorilla
Chinese hamster
Bat
|
Application | Abreviews | Notes |
---|---|---|
IHC-P | (4) |
1/1000 - 1/5000.
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ICC/IF | (6) |
Use at an assay dependent concentration.
|
WB | (10) |
1/2000 - 1/10000. Detects a band of approximately 15 kDa.
|
Flow Cyt | (1) |
Use 0.5µg for 106 cells.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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IHC-P
1/1000 - 1/5000. |
ICC/IF
Use at an assay dependent concentration. |
WB
1/2000 - 1/10000. Detects a band of approximately 15 kDa. |
Flow Cyt
Use 0.5µg for 106 cells. ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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
- 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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Samples:
Nuclear extract (50 g) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: ab11174 used at 0.1 mcg/ml. Detection: Chemiluminescence with 30 second exposure.µ UT = untreated
Ub = ubiquitylated
NCS = neocarzinostatin - 200 ng/ml, 30 minSamples: Nuclear extract (50
µg) from human HEK293, human melanoma (G361), mouse wildtype embryonic fibroblasts (+/+) or mouse H2AX knockout embryonic fibroblasts (-/-). Antibody: ab11174 used at 0.1 mcg/ml. Detection: Chemiluminescence with 30 second exposure. UT = untreated
Ub = ubiquitylated
NCS = neocarzinostatin - 200 ng/ml, 30 min -
Immunocytochemistry/Immunofluorescence analysis of neocarzinostatin treated asynchronous HeLa cells (left) and untreated asynchronous HeLa cells (right) labelling H2A.X (phospho S139 with ab11174 at 1/5000 (0.2µg/ml). A DyLight® 594-conjugated anti-rabbit IgG (1/100) was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling gamma H2A.X (phospho S139) with ab11174 at 1/5000 (0.2µg/ml). Dectection: DAB.
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ab11174 at 0.5ug staining gamma H2A.X in human Jurkat cell line by flow cytometery. Cells were treated with 5ug/ml etopside for 3 hours, fixed in 1.5% paraformaldehyde, and permeabilized in 90% methanol. A FITC-conjugated rabbit polyclonal was used as secondary (in 150ul reaction). The black line indicates cells treated with etopside and anti KLH antibody, the red line repersents untreated cells and anti gamma H2A.X and blue line indicates etopside treated anti gamma H2A.X respectively.
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Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)This image is courtesy of an anonymous AbreviewAll lanes : Anti-gamma H2A.X (phospho S139) antibody (ab11174) at 1/5000 dilution
Lane 1 : HeLa nuclear lysate - untreated
Lane 2 : HeLa nuclear lysate - IR treated
Lysates/proteins at 40 µg per lane.
Secondary
All lanes : HRP-conjugated donkey anti-rabbit IgG polyclonal
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 17 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
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Detection of gamma-H2AX by Immunofluorescence. Samples: Wildtype (H2AX +/+) or H2AX knockout (H2AX -/-) mouse embryonic fibroblasts. Antibody: Affinity purified ab11174 used at 2ug/ml. Detection: Rhodamine red labelled goat anti-rabbit IgG.
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Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174)This image is courtesy of an anonymous Abreview
ab11174 at 1/1000 staining human HeLa cells by ICC/IF. These cells express a gene that causes a DNA damage response, leading to H2AX phosphorylation. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 45 minutes. An Alexa-Fluor ® 488 conjugated goat anti-rabbit was used as the secondary.
Protocols
References (162)
ab11174 has been referenced in 162 publications.
- Ishiguro KI et al. MEIOSIN Directs the Switch from Mitosis to Meiosis in Mammalian Germ Cells. Dev Cell N/A:N/A (2020). PubMed: 32032549
- Radic M et al. The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage. Int J Mol Sci 21:N/A (2020). PubMed: 32235358
- Keenan CR et al. Extreme disruption of heterochromatin is required for accelerated hematopoietic aging. Blood 135:2049-2058 (2020). PubMed: 32305044
- Podgorski II et al. Combination of sirtuin 3 and hyperoxia diminishes tumorigenic properties of MDA-MB-231 cells. Life Sci 254:117812 (2020). PubMed: 32428596
- Polonis K et al. Chronic Intermittent Hypoxia Triggers a Senescence-like Phenotype in Human White Preadipocytes. Sci Rep 10:6846 (2020). PubMed: 32321999
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