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RabMAb

Anti-DNA PKcs抗体[Y393] (ab32566)

概述

  • 产品名称
    Anti-DNA PKcs抗体[Y393]
    参阅全部 DNA PKcs 一抗
  • 描述
    兔单克隆抗体[Y393] to DNA PKcs
  • 特异性
    This antibody is specific for DNA PKcs. It may also detect the splice isoform 2.
  • 经测试应用
    适用于: WB, IHC-P, ICC/IFmore details
    不适用于: Flow Cyt or IP
  • 种属反应性
    与反应: Mouse, Rat, Human, Armenian hamster
  • 免疫原

    corresponding to Human DNA PKcs aa 4050 to the C-terminus (C terminal).
    Database link: P78527

  • 阳性对照
    • WB: K562 cell lysate. ICC/IF: Hela cells. IHC-P: Human breast carcinoma tissue slides.
  • 常规说明

    Mouse and rat samples are recommended based on WB results.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab32566 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa).

For unpurified, use 1/1000 - 1/2000.

IHC-P 1/50.

For unpurified, use 1/5.

ICC/IF 1/100.

For unpurified, use 1/10.

  • 应用说明
    Is unsuitable for Flow Cyt or IP.
  • 靶标

    • 功能
      Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage. Involved in DNA nonhomologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination. Must be bound to DNA to express its catalytic properties. Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step. Required to protect and align broken ends of DNA. May also act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage. Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion. Also involved in modulation of transcription. Recognizes the substrate consensus sequence [ST]-Q. Phosphorylates 'Ser-139' of histone variant H2AX/H2AFX, thereby regulating DNA damage response mechanism. Phosphorylates DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, SRF, XRCC1, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2. Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA. Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D.
    • 序列相似性
      Belongs to the PI3/PI4-kinase family.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 2 HEAT repeats.
      Contains 1 PI3K/PI4K domain.
      Contains 3 TPR repeats.
    • 翻译后修饰
      Phosphorylated upon DNA damage, probably by ATM or ATR. Autophosphorylated on Thr-2609, Thr-2638 and Thr-2647. Thr-2609 is a DNA damage-inducible phosphorylation site (inducible with ionizing radiation, IR). Autophosphorylation induces a conformational change that leads to remodeling of the DNA-PK complex, requisite for efficient end processing and DNA repair.
      S-nitrosylated by GAPDH.
    • 细胞定位
      Nucleus.
    • Information by UniProt
    • 数据库链接
    • 别名
      • DNA dependent protein kinase catalytic subunit antibody
      • DNA PK catalytic subunit antibody
      • DNA-dependent protein kinase catalytic subunit antibody
      • DNA-PK catalytic subunit antibody
      • DNA-PKcs antibody
      • DNAPK antibody
      • DNAPK catalytic subunit antibody
      • DNPK 1 antibody
      • DNPK1 antibody
      • Hyper radiosensitivity of murine scid mutation, complementing 1 antibody
      • Hyperradiosensitivity complementing 1, mouse, homolog of 1 antibody
      • HYRC 1 antibody
      • HYRC antibody
      • HYRC1 antibody
      • IMD26 antibody
      • p350 antibody
      • p460 antibody
      • PKRDC antibody
      • PRKDC antibody
      • PRKDC_HUMAN antibody
      • Protein Kinase DNA Activated Catalytic Polypeptide antibody
      • XRCC 7 antibody
      • XRCC7 antibody
      see all

    Anti-DNA PKcs antibody [Y393] 图像



    • Predicted band size : 469 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: DNA PKcs knockout HAP1 cell lysate (20 µg)
      Lane 3: K562 cell lysate (20 µg)
      Lane 4: SH-SY5Y cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab32566 observed at 470 kDa. Red - loading control, ab7291, observed at 52 kDa.
      ab32566 was shown to specifically react with DNA PKCs when DNA PKcs knockout samples were used. Wild-type and ProteinX knockout samples were subjected to SDS-PAGE. ab32566 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical staining of paraffin embedded human tonsil with purified ab32566 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    • Immunofluorescent staining of HeLa cells (fixed with 4% PFA) with purified ab32566 at a dilution of 1/100. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.

    • All lanes : Anti-DNA PKcs antibody [Y393] (ab32566) at 1/7300 dilution (purified)

      Lane 1 : MOLT4 cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 469 kDa
      Observed band size : 460 kDa (why is the actual band size different from the predicted?)

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human tonsil with unpurified ab32566 at a working dilution of 1 in 5. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    • Immunofluorescent staining of HeLa cells (fixed with 4% PFA) with unpurified ab32566 at a dilution of 1/10. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.

    • All lanes : Anti-DNA PKcs antibody [Y393] (ab32566) at 1/1000 dilution (unpurified)

      Lane 1 : MOLT4 cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      HRP goat anti-rabbit (H+L) at 1/1000 dilution

      Predicted band size : 469 kDa
      Observed band size : 460 kDa (why is the actual band size different from the predicted?)

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Unpurified ab32566, at a 1/50 dilution, staining DNA PKcs in paraffin embedded human breast carcinoma tissue by immunohistochemistry.

    • Anti-DNA PKcs antibody [Y393] (ab32566) at 1/1000 dilution (unpurified) + K562 cell lysate.

      Predicted band size : 469 kDa
      Observed band size : 460 kDa (why is the actual band size different from the predicted?)

    Anti-DNA PKcs antibody [Y393] (ab32566)参考文献

    This product has been referenced in:
    • Harley ME  et al. TRAIP promotes DNA damage response during genome replication and is mutated in primordial dwarfism. Nat Genet 48:36-43 (2016). WB ; Human . Read more (PubMed: 26595769) »
    • Masuda Y  et al. TRIM29 regulates the assembly of DNA repair proteins into damaged chromatin. Nat Commun 6:7299 (2015). Human . Read more (PubMed: 26095369) »

    See all 5 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Loading amount
    20 µg
    Specification
    HeLa
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    提交于 Dec 30 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (breast cancer cell)
    Specification
    breast cancer cell
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% triton X100
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 May 09 2008

    Thank you for contacting us and your interest in our products.

    Epitomics did offer isotype controls specific for different lots of antibody purchased. This was providing the antibody at the same concentration as the antibody to be tested. We d...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Loading amount
    200000 cells
    Specification
    HEK293
    Gel Running Conditions
    Reduced Denaturing (5% gel)
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    提交于 Feb 02 2010

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (A549 tumour cell line)
    Loading amount
    50 µg
    Specification
    A549 tumour cell line
    Treatment
    100IU bleomycin for 2h
    Gel Running Conditions
    Reduced Denaturing (3-8% tris acetate gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    提交于 Nov 27 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (mouse muscle)
    Loading amount
    100 µg
    Specification
    mouse muscle
    Gel Running Conditions
    Reduced Denaturing (6% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Sep 14 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Armenian Hamster Tissue lysate - whole (Liver tissue)
    Loading amount
    50 µg
    Specification
    Liver tissue
    Gel Running Conditions
    Reduced Denaturing (6% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Sep 14 2009

    Application
    Western blot
    Sample
    African Green Monkey Cell lysate - whole cell (COS-7 cell line)
    Loading amount
    20 µg
    Specification
    COS-7 cell line
    Gel Running Conditions
    Reduced Denaturing (6% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Sep 11 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (C6 Cell line)
    Loading amount
    25 µg
    Specification
    C6 Cell line
    Gel Running Conditions
    Reduced Denaturing (6% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 11 2008

    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - nuclear (Breast cancer cells)
    Total protein in input
    200 µg
    Specification
    Breast cancer cells
    Immuno-precipitation step
    Protein A/G
    Username

    Abcam user community

    Verified customer

    提交于 May 21 2008

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