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    gamma-h2ax-phospho-s139-antibody-ab11174.pdf

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Epigenetics and Nuclear Signaling Histones Variants
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Anti-gamma H2A.X (phospho S139) 抗体 (ab11174)

  • Datasheet
  • SDS
Reviews (26) Submit a question References (231)

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Immunocytochemistry - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
  • Immunocytochemistry/ Immunofluorescence - 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
  • Reacts with: Mouse, 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

Product image
Anti-gamma H2A.X (phospho S139) antibody [EP854(2)Y] (ab81299)
  • 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

  • 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
  • 特異性

    Using IF, this antibody was shown to bind to a non-nuclear location in Hela cells.
  • アプリケーション

    適用あり: IHC-P, ICC/IF, WBmore details
  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: Rabbit, Guinea pig, Cow, Dog, Pig, Rhesus monkey, Gorilla, Chinese hamster, Bat
  • 免疫原

    Synthetic peptide corresponding to Human gamma H2A.X (phospho S139).
    Database link: 3014

  • 特記事項

    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

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • バッファー

    pH: 7
    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • 特記事項(精製)

    Antibodies were affinity purified using the peptide immobilized on solid support.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Epigenetics and Nuclear Signaling
    • Histones
    • Variants
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Human gamma H2A.X (phospho S139) peptide (ab15645)

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ab11174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
IHC-P (4)
1/1000 - 1/5000.
ICC/IF (7)
1/500 - 1/5000.
WB (11)
1/2000 - 1/10000. Detects a band of approximately 15 kDa.
特記事項
IHC-P
1/1000 - 1/5000.
ICC/IF
1/500 - 1/5000.
WB
1/2000 - 1/10000. Detects a band of approximately 15 kDa.

ターゲット情報

  • 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.
  • Target information above from: UniProt accession P16104 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 3014 Human
    • Entrez Gene: 15270 Mouse
    • Omim: 601772 Human
    • SwissProt: P16104 Human
    • SwissProt: P27661 Mouse
    • Unigene: 477879 Human
    • Unigene: 245931 Mouse
    • 別名

      • H2A histone family member X antibody
      • 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
      see all

    画像

    • Immunocytochemistry - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Immunocytochemistry - Anti-gamma H2A.X (phospho S139) antibody (ab11174)

      ab11174 staining gamma H2A.X (phospho S139) in HeLa UV cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab11174 at 0.1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      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 prostate carcinoma tissue labeling gamma H2A.X (phospho S139) with ab11174 at 1/5000 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.

    • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      All lanes : Anti-gamma H2A.X (phospho S139) antibody (ab11174) at 0.04 µg/ml

      Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) cells treated with 100 µM etoposide, whole cell lysate
      Lane 2 : NIH/3T3 cells mock treated, whole cell lysate

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG (HRP)

      Exposure time: 3 seconds


      Lower Panel: Rabbit anti-GAPDH antibody.
    • Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      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.
    • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      All lanes : Anti-gamma H2A.X (phospho S139) antibody (ab11174) at 0.04 µg/ml

      Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cells treated with 100 µM etoposide, whole cell lysate
      Lane 2 : Jurkat cells mock treated, whole cell lysate

      Lysates/proteins at 50 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG (HRP)

      Exposure time: 10 seconds


       
      Lower Panel shows western blot for total H2AX using rabbit anti-H2AX recombinant monoclonal antibody.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-gamma H2A.X (phospho S139) antibody (ab11174)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse CT26 colon carcinoma tissue labeling gamma H2A.X (phospho S139) with ab11174 at 1/5000 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.

    • Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      Western blot - Anti-gamma H2A.X (phospho S139) antibody (ab11174)This image is courtesy of an anonymous Abreview
      All 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

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-gamma H2A.X (phospho S139) antibody (ab11174)
      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.

      See Abreview

    プロトコール

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    データシートおよび資料

    • SDS download

    • Datasheet download

      Download

    参考文献 (231)

    ab11174 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab11174 は 231 報の論文で使用されています。

    • Horisawa-Takada Y  et al. Meiosis-specific ZFP541 repressor complex promotes developmental progression of meiotic prophase towards completion during mouse spermatogenesis. Nat Commun 12:3184 (2021). IHC . PubMed: 34075040
    • Cui L  et al. Mitochondrial copper depletion suppresses triple-negative breast cancer in mice. Nat Biotechnol 39:357-367 (2021). PubMed: 33077961
    • Lee JJ  et al. HMGB1 orchestrates STING-mediated senescence via TRIM30a modulation in cancer cells. Cell Death Discov 7:28 (2021). PubMed: 33558529
    • Stewart-Ornstein J  et al. p53 dynamics vary between tissues and are linked with radiation sensitivity. Nat Commun 12:898 (2021). PubMed: 33563973
    • Kato T  et al. Nuclear PTEN and p53 suppress stress-induced liver cancer through distinct mechanisms. Biochem Biophys Res Commun 549:83-90 (2021). PubMed: 33667713
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 26 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (early pachytene spermatocyte)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    early pachytene spermatocyte
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    MR. Bryan Niedenberger

    Verified customer

    投稿 Nov 16 2021

    Western blot abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (normal)
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (12)
    Loading amount
    20 µg
    Treatment
    0.4mM H2O2+FeSO4
    Specification
    normal
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 06 2021

    Immunohistochemistry (Frozen sections) abreview for Anti-gamma H2A.X (phospho S139) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (stomach organoids)
    Permeabilization
    Yes - 0.05% TX-100
    Specification
    stomach organoids
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 18 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
    Permeabilization
    No
    Specification
    Kidney
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    10% formalin
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 30 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Mesenchymal tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
    Permeabilization
    No
    Specification
    Mesenchymal tumor
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    10% formalin
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 30 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (lung)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
    Permeabilization
    No
    Specification
    lung
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    10% formalin
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 29 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (xenograft tumor)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
    Permeabilization
    No
    Specification
    xenograft tumor
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    10% formalin
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 29 2019

    Immunohistochemistry free floating abreview for Anti-gamma H2A.X (phospho S139) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry free floating
    Sample
    Rat Tissue sections (Brain sections)
    Specification
    Brain sections
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 11 2019

    Western blot abreview for Anti-gamma H2A.X (phospho S139) antibody

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (human keratinocyte cells)
    Gel Running Conditions
    Reduced Denaturing (12% SDS page gel)
    Loading amount
    10 µg
    Treatment
    30Gy irradiation for 24hr
    Specification
    human keratinocyte cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 24 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-gamma H2A.X (phospho S139) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (EpH4)
    Permeabilization
    Yes - 0.2% triton
    Specification
    EpH4
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 16°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 08 2018

    1-10 of 26 Abreviews or Q&A

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