Phospho-Histone H2A.X (Ser139)[Ser140] Antibody

製品:	Phospho-Histone H2A.X (Ser139)[Ser140] Antibody
カタログ:	AF3187
タンパク質の説明:	Rabbit polyclonal antibody to Phospho-Histone H2A.X (Ser139)[Ser140]
アプリケーション:	WB IHC
Cited expt.:	WB, IHC
反応性:	Human, Mouse, Rat
予測:	Bovine, Sheep, Dog
分子量:	15kDa; 15kD(Calculated).
ユニプロット:	P16104
RRID:	AB_2834619

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サイズ	価格	在庫状況
100ul	$280	在庫あり
200ul	$350	在庫あり

プロトコル

製品説明

ソース:

Rabbit

アプリケーション:

WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*Tips:

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.

反応性:

Human,Mouse,Rat

予測:

Bovine(91%), Sheep(91%), Dog(91%)

クローナリティ:

Polyclonal

特異性:

Phospho-Histone H2A.X (Ser139) Antibody detects endogenous levels of Histone H2A.X only when phosphorylated at Ser140, which site historically referenced as Ser139.

RRID:

AB_2834619
引用形式: Affinity Biosciences Cat# AF3187, RRID:AB_2834619.

コンジュゲート:

Unconjugated.

精製:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

保存:

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.

別名:

折りたたみ／展開

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;

免疫原

免疫原:

A synthesized peptide derived from human Histone H2A.X around the phosphorylation site of Ser139.

Uniprot:

P16104(H2AX_HUMAN) >>Visit HPA database.

遺伝子(ID):

H2AFX(3014)

タンパク質の説明:

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.

タンパク質配列:

P16104 H2AX_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MSGRGKTGGKARAKAKSRSSRAGLQFPVGRVHRLLRKGHYAERVGAGAPVYLAAVLEYLTAEILELAGNAARDNKKTRIIPRHLQLAIRNDEELNKLLGGVTIAQGGVLPNIQAVLLPKKTSATVGPKAPSGGKKATQASQEY

種類予測

種類予測:

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.

Species

Results

Score

Bovine

Sheep

Dog

Pig

Horse

Xenopus

Zebrafish

Chicken

Rabbit

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

研究背景

P16104_H2AX_HUMAN

機能:

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.

PTMs:

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).

細胞の位置付け:

Nucleus. Chromosome.

タンパク質ファミリー:

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

Belongs to the histone H2A family.

研究領域

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

· Human Diseases > Substance dependence > Alcoholism.

· Human Diseases > Immune diseases > Systemic lupus erythematosus.

参考文献

1). Intracellular Self-Assembled Fe(II)-Aniline Mustard NPs for Self-Enhanced Anti-Tumor Chemo-/Chemodynamic Therapy. ADVANCED FUNCTIONAL MATERIALS, 2025 [IF=19.0]

2). NFS1, together with FXN, protects cells from ferroptosis and DNA damage in diffuse large B-cell lymphoma. Redox biology, 2025 (PubMed: 41005206) [IF=10.7]

Application: WB Species: human Sample: DLBCL cells

Fig. 7. ISC proteins promote DNA damage repair in DLBCL cells (A-B) Cell cycle distribution in ISC proteins downregulated cells. (C–D) Protein expression of DNA repair-related proteins CHK1, pCHK1(S345), H2AX, and pH2AX(S139) in ISC proteins downregulated cells. (E–F) The protein expression of DNA polymerases POLA1, POLD1, and POLE in ISC proteins downregulated cells. Data were expressed as mean ± SD (N = 3). KD-NC means negative control, and NFS1-KD or FXN-KD means the knockdown of NFS1 or FXN. ∗p < 0.05 NFS1-KD vs. KD-NC, ∗∗p < 0.01 NFS1-KD vs. KD-NC, ##p < 0.01 FXN-KD vs. KD-NC.

3). Exposure to Sucralose and Its Effects on Testicular Damage and Male Infertility: Insights into Oxidative Stress and Autophagy. Environmental health perspectives, 2025 (PubMed: 40378307) [IF=10.1]

4). BM-MSC-derived migrasomes reverse stroke-induced thymic atrophy and immunosuppression via Pin1 delivery to thymic epithelial cells. Journal of neuroinflammation, 2025 (PubMed: 41241718) [IF=9.3]

Application: WB Species: Mouse Sample:

Fig. 2 BM-MSC transplantation promotes the proliferation of medullary thymic epithelial cells (mTEC). Male wild-type C57/Bl6 mice underwent 60 minutes of tMCAO followed by reperfusion. BM-MSC (2 × 10⁶ cells/mouse) or vehicle (Veh) were administered intravenously as a single dose 2 hours postreperfusion. Thymic tissues were harvested at 7 days post-tMCAO. A-B Bulk RNA sequencing (RNA-seq) analysis of thymic tissues at 7 days post-tMCAO (n = 4 per group). A Volcano plot of differentially expressed genes (DEGs) between the BM-MSC and Veh groups. B Gene Ontology (GO) enrichment analysis of the top 20 enriched biological processes (BP) among the DEG. C Western blot analysis of proliferation-related (Ki67) and senescence-related (NAMPT, p21, and γH2AX) protein expression. The bar graphs show the relative protein levels (n = 4 per group). *p

5). Novel intravenous formulation for radiosensitization in osteosarcoma treatment. Materials today. Bio, 2025 (PubMed: 40206141) [IF=8.7]

Application: WB Species: Mouse Sample: K7M2 cells

Fig. 3. Anti-OS effects of HPTM combined with NIR and RT. (A) Fluorescence microscopy images and (D) quantitative analysis of ROS in K7M2 cells which were stained with DCFH-DA after different treatments, including HPTM, HP + NIR + RT, RT, HPTM + NIR, HPTM + RT, HPTM + NIR + RT. (B) Live/dead cell staining was performed through Calcein AM (green)/PI (red) double staining method. (C, E) Evaluation of apoptosis rates of K7M2 cells treated with or without X-ray (4 Gy) in the presence or not the presence of NIR for 48h. (F) Cell viability was assessed by CCK8 method. (G) Expression of pro-apoptotic proteins Bax, anti-apoptotic proteins Bcl-2 and DNA double-strand breaks marker γH2A.x in K7M2RR OS cells after different treatment. All experiments were carried out with 3 biological replicates. # indicates a group which was compared with other groups. All values are the mean ± SD. P values, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001,∗∗∗∗p < 0.0001.

6). miR-211 facilitates platinum chemosensitivity by blocking the DNA damage response (DDR) in ovarian cancer. Cell Death & Disease, 2019 (PubMed: 31235732) [IF=8.1]

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.

7). Reduction of DNA Topoisomerase Top2 Reprograms the Epigenetic Landscape and Extends Health and Life Span Across Species. Aging cell, 2025 (PubMed: 39935344) [IF=8.0]

Application: WB Species: human Sample: IMR-90 cells

FIGURE 4 TOP2B knock down reduces various cellular aging hallmarks in human IMR-90 cells and in various mouse tissues. (A–H) IMR-90 cells were induced to undergo replicative, stress-induced, or oncogene-induced senescence. The protein levels of p16, p21, and γH2AX were detected by western blotting (A, B). γH2AX foci in DAPI-stained nuclei of IMR-90 cells were measured by immunofluorescent staining (C, D). The protein level of PARP1 was detected by western blotting (E, F). SA-β-Gal staining (blue-stained cells) and quantification of percent of SA-β-Gal positive cells were shown in G, H. (I–P) Confocal fluorescence images of IMR-90 cells (I–K) and C. elegans (N–P) stained by LysoTracker Red DND-99 (LTR) and LysoSensorTM Green DND-189 (LSG), and the fluorescence intensity ratio of LSG/LTR was measured as an indicator of lysosomal acidity. (L, M) IMR-90 cells were induced to undergo replicative, stress-induced, or oncogene-induced senescence. The protein level of TFEB was detected by western blotting. The relative intensity of LSG/LTR and average lysosome diameter in IMR-90 cells (J, K) and C. elegans (O, P) were quantified. (Q, R) Western blot analysis of nutrient-sensing mTOR signaling proteins in IMR-90 cells. (S–Y) Western blot analysis of nutrient-sensing mTOR signaling proteins in mouse kidney, lung, and skeletal muscle. Statistical analysis was performed using GraphPad Prism v8.0 software (https://www.graphpad.com). Data were considered statistically significant at p

8). FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination. Clinical and translational medicine, 2022 (PubMed: 35858239) [IF=7.9]

Application: IF/ICC Species: human Sample:

FIGURE 1 Oocyte-predominant FBXW24 is essential for female fertility. (A) Q-PCR shows that Fbxw24 mRNA was predominant in ovaries. (B and C) Western blots and quantification demonstrate that FBXW24 is more dominant in oocytes than in granular cells (GCs). (D and E) Western blots and quantification show that FBXW24 protein peaked at 16.5 DPC and then gradually decreased from PND 1 to 21. (F–H) Five bases from the third exon of the Fbxw24 gene were deleted through Cas9 and caused a frame-shift of the Fbxw24 gene; western blots show that in Fbxw24-KO ovaries, FBXW24 protein is completely eliminated. (I–K) At PND 21, Fbxw24 knockout significantly increased primordial follicles (PMF) while it decreased primary follicles (PF), secondary follicles (SF), and antral follicles (AF). Four selected regions (red dot-line square) from WT and KO ovaries were zoomed and placed on the right, and primordial follicles were arrow-pointed. (L) Curves for cumulative pups from 2-month-old to 7-month-old showed that Fbxw24-KO male mice had normal fertility, while Fbxw24-KO female mice were completely infertile. (M and N) Fbxw24 knockout significantly decreased oocyte maturation rate (first polar body, 1 pb). (O and P) Fbxw24 knockout significantly decreased numbers of ovulated oocytes. (Q and R) Immunofluorescence and quantification showed that Fbxw24 knockout significantly increased DSBs (by γH2AX foci) in the GV oocytes. DNA in blue, γH2AX in green. (S) Live imaging shows that oocytes were not able to go through anaphase because homologous chromosomes did not separate at all. Red fluorescence signals are H2B, and green fluorescence signals are α-Tubulin. Scale bar in Q, 20 μm; Scale bar in other panels, 100 μm. GAPDH was used as a loading control. *, p < 0.05; **, p < 0.01; ****, p < 0.0001. AU, arbitrary unit

9). Danhong injection alleviates cerebral ischemia/reperfusion injury by improving intracellular energy metabolism coupling in the ischemic penumbra. Biomedicine & Pharmacotherapy, 2021 (PubMed: 34058441) [IF=6.9]

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.

10). Stilbene B10 induces apoptosis and tumor suppression in lymphoid Raji cells by BTK-mediated regulation of the KRAS/HDAC1/EP300/PEBP1 axis. Biomedicine & Pharmacotherapy, 2022 (PubMed: 36274467) [IF=6.9]

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Phospho-Histone H2A.X (Ser139)[Ser140] Antibody - #AF3187

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