Overview

  • Product nameAnti-DNA PKcs antibodySee all DNA PKcs primary antibodies ...
  • Description
    Rabbit polyclonal to DNA PKcs
  • Tested applicationsIP, WB, ICC/IF more details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 3650 - 3750 of Human DNA PKcs.

    (Peptide available as ab41959.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: HeLa Nuclear, MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell and SW480 (Human colon adenocarcinoma cell line) Whole Cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab39978 in the following tested applications.

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

Application Notes
IP Use at an assay dependent concentration.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa).Can be blocked with Human DNA PKcs peptide (ab41959).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionSerine/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 similaritiesBelongs 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 localizationNucleus.
  • Target information above from: UniProt accession P78527 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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
    • HYRC 1 antibody
    • HYRC antibody
    • HYRC1 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 images

  • All lanes : Anti-DNA PKcs antibody (ab39978) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 3 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
    Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate (ab3957)

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 469 kDa
    Observed band size : 460 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab39978 stained HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab39978, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue). This antibody also gave a positive IF result in HepG2 cells fixed in 4% PFA at 1ug/ml. However, this Fast-Track antibody is not yet fully characterised. This image represents inconclusive preliminary data.
  • DNA PKcs was immunoprecipitated using 0.5mg SW480 whole cell extract, 5µg of Rabbit polyclonal to DNA PKcs and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, SW480 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab39978.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Band: 460kDa: DNA PKcs.

References for Anti-DNA PKcs antibody (ab39978)

ab39978 has not yet been referenced specifically in any publications.

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