Overview

  • Product name

    Anti-DNA PKcs antibody [EPR392]
    See all DNA PKcs primary antibodies
  • Description

    Rabbit monoclonal [EPR392] to DNA PKcs
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt,IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human DNA PKcs aa 50-150. The exact sequence is proprietary.
    Database link: P78527

  • Positive control

    • WB: K562, Molt4, MCF7, SH-SY5Y, 293T and PC3 cell lysates. ICC/IF: K-562 cells
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab133516 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/10000. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa).
ICC/IF 1/200.

For unpurified use at 1/500 - 1/1000.

  • Application notes
    Is unsuitable for Flow Cyt,IHC-P or IP.
  • Target

    • Function

      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.
    • Sequence similarities

      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.
    • Post-translational
      modifications

      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.
    • Cellular localization

      Nucleus.
    • Information by UniProt
    • Database links

    • Alternative names

      • 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

    Images

    • Lane 1: Wild type HAP1 whole cell lysate (40 µg)
      Lane 2: DNA PKcs knockout HAP1 whole cell lysate (40 µg)
      Lane 3: K562 whole cell lysate (20 µg)
      Lane 4: SHSY5Y whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab133516 observed at 460 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab133516 was shown to specifically react with DNA PKcs when DNA PKcs knockout samples were used. Wild-type and DNA PKcs knockout samples were subjected to SDS-PAGE. Ab133516 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

       

    • All lanes : Anti-DNA PKcs antibody [EPR392] (ab133516) at 1/5000 dilution (Purified)

      Lane 1 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
      Lane 2 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

    • Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling DNA PKcs with purified ab133516 at 1:200 dilution (9.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • All lanes : Anti-DNA PKcs antibody [EPR392] (ab133516) at 1/1000 dilution

      Lane 1 : K562 cell lysate
      Lane 2 : Molt4 cell lysate
      Lane 3 : MCF7 cell lysate
      Lane 4 : SH-SY5Y cell lysate
      Lane 5 : 293T cell lysate
      Lane 6 : PC3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

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

    References

    This product has been referenced in:

    • Fujiwara Y  et al. A Nucleolar Stress-Specific p53-miR-101 Molecular Circuit Functions as an Intrinsic Tumor-Suppressor Network. EBioMedicine 33:33-48 (2018). Read more (PubMed: 30049386) »
    See 1 Publication for this product

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