Anti-gamma H2A.X (phospho S139) 抗体 (ab2893)
Key features and details
- Rabbit polyclonal to gamma H2A.X (phospho S139)
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
リコンビナント抗体で、ロット間での高い再現性を実現
- 異なるロット間での安定した再現性
- 容易なスケールアップ
- 評価試験による特異性の確認済み
- 倫理基準に準拠 - アニマル・フリーの生産
製品の概要
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製品名
Anti-gamma H2A.X (phospho S139) antibody
gamma H2A.X 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to gamma H2A.X (phospho S139) -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, WBmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Rat, Chimpanzee -
免疫原
Synthetic peptide conjugated to KLH derived from within residues 100 - 200 of Human gamma H2A.X, phosphorylated at S139.
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特記事項
ab2893 is batch tested in peptide array, western blot and ICC only, although some customers have successfully used this product in IHC and ChIP (see images below). We would recommend ab81299 as an alternative product for use in IHC and ChIP.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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ChIP Related Products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Camptothecin, DNA topoisomerase inhibitor (ab120115)
- Terfenadine, K+ channel blocker. H1 antagonist. (ab120270)
- TMPyP4 tosylate, Telomerase inhibitor (ab120793)
- 10-Hydroxycamptothecin, DNA topoisomerase I inhibitor (ab141071)
- CPT 11 (Irinotecan), DNA topoisomerase I inhibitor (ab141107)
- SN 38, DNA topoisomerase I inhibitor (ab141108)
- Anti-BRCA1 antibody [MS110] (ab16780)
- Anti-ATM (phospho S1981) antibody [10H11.E12] (ab36810)
- Anti-Histone H2A.Z antibody [4A4] (ab80150)
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab2893の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | (7) |
Use a concentration of 1 - 5 µg/ml.
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WB | (10) |
1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa).
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特記事項 |
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ICC/IF
Use a concentration of 1 - 5 µg/ml. |
WB
1/1000. Detects a band of approximately 17 kDa (predicted molecular weight: 17 kDa). |
ターゲット情報
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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. -
配列類似性
Belongs to the histone H2A family. -
発生段階
Synthesized in G1 as well as in S-phase. -
ドメイン
The [ST]-Q motif constitutes a recognition sequence for kinases from the PI3/PI4-kinase family. -
翻訳後修飾
Phosphorylated 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. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
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参照データベース
- 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
see all -
別名
- H2A histone family member X antibody
- H2A histone family member X antibody
- H2A.FX antibody
see all
画像
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Lanes 1-2 : Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1/500 dilution
Lanes 3-4 : Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1/500 dilution (+ Histone H2A.X (phospho Ser 139) peptide ab15645)
Lanes 5-6 : Anti-gamma H2A.X (phospho S139) antibody (ab2893) at 1/500 dilution (+ Histone H2A.X non-phospho peptide - ab15646)
Lanes 1 & 3 & 5 : Control HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate Histone preparation
Lanes 2 & 4 & 6 : Colcemid treated HeLa whole cell lysate Histone preparation
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution (Goat polyclonal to Rabbit IgG (HRP))
Predicted band size: 17 kDaBlocking peptides at 1ug/lane.
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Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)Garcia, C.P. et al PLoS One. 2016; 11(3): e0152607. Fig.2a Published online 2016 Mar 31. doi: 10.1371/journal.pone.0152607 Reproduced under the Creative Commons licence https://creativecommons.org/licenses/by/4.0/
Immunofluorescence photomicrographs of genotoxic-treated (1μM during 3 h) human induced pluripotent stem cells (hiPSCs) and hESCs-derived neuroprogenitors (NP) performed immediately after CPT treatment (1μM during 3 h). The figure shows representative images of cells stained with primary antibodies against ATM phospho-serine1981 (pATM), histoneγH2AX, p53, p53 phospho serine 15 (p53pSer15). Nuclei were counterstained with DAPI. The scale bars represent 100 μm.
Histone gamma H2A.X was detected using ab2893.
From Figure 2a of Garcia et al PLoS One. 2016; 11(3): e0152607. Published online 2016 Mar 31. doi: 10.1371/journal.pone.0152607
Reproduced under the Creative Commons licence: https://creativecommons.org/licenses/by/4.0/
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab2893)Image from Dr CD Curtis et al, BMC Cancer. 2010 Jan 11;10:9, Fig 11.
ab2893 staining gamma H2A.X in human mammary tissue by Immunohistochemistry (paraffin embedded sections).Paraffin-embedded blocks were sectioned and mounted on frost-free slides. The 3-10 µm sections were deparaffinized in xylene and rehydrated through a series of graded alcohols. Slides were washed with 1× PBS and endogenous peroxidases were blocked with 1.5% hydrogen peroxide in 1× PBS for 20 minutes at 25°C. After three 5 minutes washes in 1× PBS, slides were incubated in blocking solution (1× PBS with 0.1% Triton X-100, 3% bovine serum albumin) with 5% normal donkey serum for 10 minutes at 25°C. Control (no primary antibody) and experimental slides were incubated overnight at 4°C, respectively, in blocking solution alone or blocking solution with ab2893 at 1/1000 dilution. Biotin-conjugated secondary antibody 1/200 was added and slides were incubated at 25°C for 30 minutes and then washed three times with 1× PBS. The ABC Peroxidase Staining kit (1/100 dilution of each Reagent A and B in 1× PBS) was applied at 25°C for 30 min. After 3 washes with 1× PBS, staining was visualized with peroxidase-sensitive Sigmafast 3,3'-Diaminobenzidine tablets. All slides were counterstained with 0.1% methyl green for 3 min at 60°C, dehydrated in ethanol, cleared in xylene and mounted with Permount. Images were obtained at 40× using a Leica DMI4000B confocal microscope with the Retiga 2000R digital camera. Exposure times were kept constant for all samples.
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ICC/IF image of ab2893 stained UV treated HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were 100% methanol fixed (5 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab2893 at 1 µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43 µM for 1 hour at room temperature.
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Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)This image is courtesy of Kirk McManus
Asynchronous HeLa cells were paraformaldehyde fixed and immunofluorescently labeled with ab2893 that had been preincubated with either 1) non-phosphorylated or 2) phosphorylated H2AX peptide. Identical exposure times were employed. The Merge images present the DAPI and ab2893 channels as red and green, respectively. Scale bars represent 5
m.µ 1) Non-phosphorylated peptides
2) Phosphorylated peptides
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HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were incubated at 37°C for 3h with vehicle control (0 µM) and different concentrations of camptothecin (ab120115). Increased expression of γH2A.X (phospho S139) in HeLa cells correlates with an increase in camptothecin concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 20µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab2893 at 1 µg/ml and ab10475 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.
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ab2893 staining γH2A.X in MALME-3M cells treated with terfenadine (ab120270), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of terfenadine, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of ab120270 (terfenadine) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (10 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab2893 staining γH2AX (phospho S139) in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with camptothecin (ab120115), by ICC/IF. Increased nuclear expression of γH2AX (phospho S139) correlates with increased concentration of camptothecin, as described in literature.
The cells were incubated at 37°C for 3h in media containing different concentrations of ab120115 (camptothecin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with SN 38 (ab141108), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of SN 38, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of ab141108 (SN 38) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with CPT 11 (Irinotecan) (ab141107), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of CPT 11 (Irinotecan), as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of ab141107 (CPT 11 (Irinotecan)) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab2893 staining γH2A.X in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 10-Hydroxycamptothecin (ab141071), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of 10-Hydroxycamptothecin, as described in literature.
The cells were incubated at 37°C for 6 hours in media containing different concentrations of ab141071 (10-Hydroxycamptothecin) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
ab2893 staining γH2A.X in weri cells treated with TMPyP4 tosylate (ab120793), by ICC/IF. Increase of γH2A.X nuclear expression correlates with increased concentration of TMPyP4 tosylate, as described in literature.
The cells were incubated at 37°C for 24 hours in media containing different concentrations of ab120793 (TMPyP4 tosylate ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2893 (1 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue. -
Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab2893)Image courtesy of Dr. Kirk McManus
Asynchronous HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were exposed to 2Gy and permitted to recover for 30min. Cells were paraformaldehyde fixed (4%), immunofluorescently labeled with ab2893 and counterstained with DAPI. The merge image presents the DAPI and ab2893 channels as red and green, respectively. The scale bar represents 5µm.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (269)
ab2893 は 269 報の論文で使用されています。
- Orgaz JL et al. Myosin II Reactivation and Cytoskeletal Remodeling as a Hallmark and a Vulnerability in Melanoma Therapy Resistance. Cancer Cell 37:85-103.e9 (2020). PubMed: 31935375
- Yin N et al. IDH1-R132H mutation radiosensitizes U87MG glioma cells via epigenetic downregulation of TIGAR. Oncol Lett 19:1322-1330 (2020). PubMed: 31966064
- Wakita M et al. A BET family protein degrader provokes senolysis by targeting NHEJ and autophagy in senescent cells. Nat Commun 11:1935 (2020). PubMed: 32321921
- Shao Z et al. CPNE1 predicts poor prognosis and promotes tumorigenesis and radioresistance via the AKT singling pathway in triple-negative breast cancer. Mol Carcinog 59:533-544 (2020). PubMed: 32181526
- Tan L et al. Interferon regulatory factor-1 suppresses DNA damage response and reverses chemotherapy resistance by downregulating the expression of RAD51 in gastric cancer. Am J Cancer Res 10:1255-1270 (2020). PubMed: 32368400
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