Product: Phospho-Histone H2A.X (Ser139)[Ser140] Antibody
Catalog: AF3187
Description: Rabbit polyclonal antibody to Phospho-Histone H2A.X (Ser139)[Ser140]
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Bovine, Sheep, Dog
Mol.Wt.: 15kDa; 15kD(Calculated).
Uniprot: P16104
RRID: AB_2834619

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 100ul $280 In stock
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Product Info

WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Bovine(91%), Sheep(91%), Dog(91%)
Phospho-Histone H2A.X (Ser139) Antibody detects endogenous levels of Histone H2A.X only when phosphorylated at Ser140, which site historically referenced as Ser139.
Cite Format: Affinity Biosciences Cat# AF3187, RRID:AB_2834619.
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.


AW228881; H2A histone family member X; H2A.FX; H2A.X; H2a/x; H2AFX; H2AX; H2AX histone; H2AX_HUMAN; Hist5.2ax; Histone 2A; Histone 2AX; Histone H2A.X; Histone H2AX; RGD1566119; γH2AX;gamma-H2AX;γ-H2AX;


Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes.



Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P16104 As Substrate

Site PTM Type Enzyme
S2 Phosphorylation
K6 Acetylation
K10 Acetylation
K14 Ubiquitination
K16 Ubiquitination
S19 Phosphorylation
K119 Acetylation
K119 Ubiquitination
K120 Ubiquitination
T121 Phosphorylation
S122 Phosphorylation
T124 Phosphorylation
K134 Methylation
K134 Sumoylation
K134 Ubiquitination
T137 Phosphorylation
S140 Phosphorylation Q13315 (ATM) , P45984 (MAPK9) , P45983 (MAPK8)
Y143 Phosphorylation Q9UIG0 (BAZ1B)

Research Backgrounds


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.


Phosphorylated on Ser-140 (to form gamma-H2AX 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 (By similarity). 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. Ubiquitination at Lys-14 and Lys-16 (H2AK13Ub and H2AK15Ub, respectively) in response to DNA damage is initiated by RNF168 that mediates monoubiquitination at these 2 sites, and 'Lys-63'-linked ubiquitin are then conjugated to monoubiquitin; RNF8 is able to extend 'Lys-63'-linked ubiquitin chains in vitro. H2AK119Ub and ionizing radiation-induced 'Lys-63'-linked ubiquitination (H2AK13Ub and H2AK15Ub) are distinct events.

Acetylation at Lys-37 increases in S and G2 phases. This modification has been proposed to play a role in DNA double-strand break repair (By similarity).

Subcellular Location:

Nucleus. Chromosome.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Subunit Structure:

The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA (Probable). Interacts with numerous proteins required for DNA damage signaling and repair when phosphorylated on Ser-140. These include MDC1, TP53BP1, BRCA1 and the MRN complex, composed of MRE11, RAD50, and NBN. Interaction with the MRN complex is mediated at least in part by NBN. Also interacts with DHX9/NDHII when phosphorylated on Ser-140 and MCPH1 when phosphorylated at Ser-140 or Tyr-143. Interacts with ARRB2; the interaction is detected in the nucleus upon OR1D2 stimulation. Interacts with WRAP53/TCAB1.

(Microbial infection) Interacts with Epstein-Barr virus protein EBNA6.


The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family.

Belongs to the histone H2A family.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Human Diseases > Substance dependence > Alcoholism.

· Human Diseases > Immune diseases > Systemic lupus erythematosus.


1). miR-211 facilitates platinum chemosensitivity by blocking the DNA damage response (DDR) in ovarian cancer. Cell Death & Disease (PubMed: 31235732) [IF=9.0]

Application: WB    Species: human    Sample: HO8910 and SKOV3 cells

Fig. 4 |TDP1 is the key target mediating the function of miR-211. a, c The shRNA technique effectively blocked the expression of TDP1 in HO8910 and SKOV3 cells. b, d Downregulation of TDP1 facilitated the sensitivity of HO8910 and SKOV3 cells to carboplatin. e Knockdown of TDP1 resulted in the upregulation of γ-H2AX in HO8910 and SKOV3 cells treated with carboplatin.

2). Danhong injection alleviates cerebral ischemia/reperfusion injury by improving intracellular energy metabolism coupling in the ischemic penumbra. Biomedicine & Pharmacotherapy (PubMed: 34058441) [IF=7.5]

Application: WB    Species: rat    Sample:

Fig. 3. |Effect of DHI on oxidative stress and DNA injure in the ischemic penumbra.D-G: Representative images of WB analysis and the semi-quantification of SOD2, RhoGDI and γ-H2A.X. Data are expressed as mean ± SD (n = 3).

Application: IHC    Species: rat    Sample: brain

Fig. 3. |Effect of DHI on oxidative stress and DNA injure in the ischemic penumbra.H and I: Immunostaining photomicrographs of γ-H2A.X and quantitative analysis of the IOD. Bar = 50 µm. Data are expressed as mean ± SD (n = 4). *P < 0.05 versus CI/R group; **P < 0.01 versus CI/R group; ##P < 0.01 versus sham group.

3). Stilbene B10 induces apoptosis and tumor suppression in lymphoid Raji cells by BTK-mediated regulation of the KRAS/HDAC1/EP300/PEBP1 axis. Biomedicine & Pharmacotherapy (PubMed: 36274467) [IF=7.5]

4). The activated ATM/p53 pathway promotes autophagy in response to oxidative stress-mediated DNA damage induced by Microcystin-LR in male germ cells. Ecotoxicology and Environmental Safety (PubMed: 34715501) [IF=6.8]

Application: WB    Species: Mouse    Sample: GC-1 cells

Fig. 2. The condition of DNA damage in GC-1 cells and mouse testis. (A) Detection of DNA damage by comet assay and the statistics of OTM and TailDNA% in GC-1 cells. Bar= 50 µm. (B and C) The expression level of γ-H2AX protein in testicular tissues and GC-1 cells was detected with Western Blot. *p < 0.05 vs. the control group; #p < 0.05 vs. the corresponding MC-LR exposure group. All data were expressed as  ± SD (n = 3).

5). Microcystin-LR accelerates follicular atresia in mice via JNK-mediated adherent junction damage of ovarian granulosa cells. Ecotoxicology and Environmental Safety (PubMed: 36731181) [IF=6.8]

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