Anti-gamma H2A.X (phospho S139) antibody (ab11174)
- Product nameAnti-gamma H2A.X (phospho S139) antibodySee all gamma H2A.X primary antibodies ...
- DescriptionRabbit polyclonal to gamma H2A.X (phospho S139)
- SpecificityUsing IF, this antibody was shown to bind to a non-nuclear location in Hela cells.
- Tested applicationsICC/IF, WB, Flow Cyt, ICC more details
- Species reactivityReacts with: Mouse, Chicken, Human
Predicted to work with: Rabbit, Guinea pig, Cow, Dog, Pig, Rhesus monkey, Gorilla, Chinese Hamster, Bat
Synthetic phospho-peptide, which represents a portion of the C-terminus of human histone H2AX surrounding phosphorylated serine 139 (LocusLink ID 3014).
- Positive control
- Tested with human HEK293, human G-361 and mouse embryonic fibroblast cells.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage bufferPreservative: 0.1% Sodium Azide
Constituents: 8mM PBS, 60mM Citrate, 150mM Tris, pH 7-8
- Concentration information loading...
- PurityImmunogen affinity purified
- Purification notesAntibodies were affinity purified using the peptide immobilized on solid support.
- Clonality Polyclonal
- Research Areas
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.
|ICC/IF||ICC/IF: Use at an assay dependent dilution.|
|WB||WB: 1/5000 - 1/15000. Detects a band of approximately 15 kDa.|
|Flow Cyt||Flow Cyt: Use 0.5µg for 106 cells.|
|ICC||ICC: 1/400 - 1/800.|
- FunctionVariant 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 similaritiesBelongs to the histone H2A family.
- Developmental stageSynthesized in G1 as well as in S-phase.
- DomainThe [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.
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 localizationNucleus. Chromosome.
- H2A histone family member X antibody
- H2A.FX antibody
- H2A.X antibody
- H2A/X antibody
- H2AFX antibody
- H2AX antibody
- H2AX_HUMAN antibody
- Histone H2A.x antibody
Anti-gamma H2A.X (phospho S139) antibody images
Samples: 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
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.
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.
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.
References for Anti-gamma H2A.X (phospho S139) antibody (ab11174)
This product has been referenced in:
- Ivanov A et al. Lysosome-mediated processing of chromatin in senescence. J Cell Biol 202:129-43 (2013). Read more (PubMed: 23816621) »
- Zhang X et al. Diaminothiazoles modify Tau phosphorylation and improve the tauopathy in mouse models. J Biol Chem 288:22042-56 (2013). Read more (PubMed: 23737518) »