Overview

  • Product name
  • Description
    Rabbit polyclonal to DNA PKcs
  • Host species
    Rabbit
  • Specificity
    Ku-80 co-immunoprecipitates using this antibody.
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Chicken, Guinea pig, Cow, Pig, Chimpanzee, Rhesus monkey, Gorilla, Chinese hamster, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human DNA PKcs (C terminal). Immunogen is in the region of aa 4075 and the c terminus.
    Database link: P78527

  • Positive control
    • Whole cell lysate from Hela cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab70250 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 469 kDa (predicted molecular weight: 469 kDa).
IP Use at an assay dependent concentration.

Use at 2 to 5 µg/mg lysate.

ICC/IF Use at an assay dependent concentration.

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

  • All lanes : Anti-DNA PKcs antibody (ab70250) at 0.1 µg/ml

    Lane 1 : Whole cell lysate from Hela cells at 15 µg
    Lane 2 : Whole cell lysate from Hela cells at 50 µg

    Predicted band size: 469 kDa
    Observed band size: 469 kDa
    Additional bands at: 170 kDa, 180 kDa, 230 kDa. We are unsure as to the identity of these extra bands.

  • ab70250 staining DNA PKCs in T98G Human brain glioblastoma by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100, pH 7.4 for 5 minutes at room temperature and blocked with 5% BSA for 10 minutes at room temperature. Samples were incubated with primary antibody (1/200 in PBS) for 1 hour. A CF568-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • Immunoprecipitation/ Western Blot of DNA PKcs
    Lane 1: ab70250 at 3µg/100µg for IP of HeLa whole cell lysate (1mg) followed by ab70250 at 0.1µg/ml for WB.
    Lane 2: Control IgG.

    Chemiluminescence with an exposure time of 30 seconds.

References

This product has been referenced in:
  • Sekiba K  et al. DHX9 regulates production of hepatitis B virus-derived circular RNA and viral protein levels. Oncotarget 9:20953-20964 (2018). Read more (PubMed: 29765512) »
  • Simões-Sousa S  et al. The p38a Stress Kinase Suppresses Aneuploidy Tolerance by Inhibiting Hif-1a. Cell Rep 25:749-760.e6 (2018). Read more (PubMed: 30332653) »
See all 9 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Abreviews
Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (CHO AA8, CHO Xrs6, CHO V-3)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
50 µg
Specification
CHO AA8, CHO Xrs6, CHO V-3

Doris Niewolik

Verified customer

Submitted Nov 28 2017

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: RT°C
Sample
Human Cell (T98G Human brain glioblastoma)
Specification
T98G Human brain glioblastoma
Permeabilization
Yes - 0.1% v/v Triton X-100 pH 7.4 for 5 min at RT
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Dec 12 2013

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