Overview

  • Product nameAnti-DNA PKcs antibody [18-2]
    See all DNA PKcs primary antibodies
  • Description
    Mouse monoclonal [18-2] to DNA PKcs
  • SpecificityThis antibody is specific to 460 kDa protein known as DNA-dependent Protein Kinase (DNA-PKcs).
  • Tested applicationsFunctional Studies, IHC-P, IHC-Fr, IP, WB, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Human DNA-dependent Protein Kinase purified from HeLa cells.

  • EpitopeAmino acids 1-2713.
  • Positive control
    • LS174T cells. Tonsil.

Properties

Applications

Our Abpromise guarantee covers the use of ab1832 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
Functional Studies Use at an assay dependent dilution.
IHC-P Use a concentration of 2 - 4 µg/ml. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0,for 10-20 min followed by cooling at RT for 20 min.
IHC-Fr Use at an assay dependent dilution.
IP Use at an assay dependent dilution. (protein G) Use at a concentration of 2µg/mg of protein lysate (native and denatured).
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa). This antibody reacts with intact DNA-PKcs as well as its degradation products.
ICC/IF Use at an assay dependent dilution. PubMed: 20709753
Flow Cyt Use at an assay dependent concentration.

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.
  • 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
    • DNPK1 antibody
    • Hyper radiosensitivity of murine scid mutation, complementing 1 antibody
    • HYRC 1 antibody
    • HYRC antibody
    • HYRC1 antibody
    • HYRC1 antibody
    • p350 antibody
    • p460 antibody
    • PKRDC antibody
    • PRKDC antibody
    • PRKDC_HUMAN antibody
    • Protein Kinase DNA Activated Catalytic Polypeptide antibody
    • Protein Kinase DNA Activated Catalytic Polypeptide antibody
    • XRCC 7 antibody
    • XRCC7 antibody
    • XRCC7 antibody
    see all

Anti-DNA PKcs antibody [18-2] images

  • ab1832 staining DNA PKcs in HCT 116 cells by Immunocytochemistry/ Immunofluorescence.
    The cells were washed in PBS, fixed in 4% paraformaldehyde (in PBS) for 15 minutes, permeabilized in 0.3% Triton X-100 at room temperature for 10 minutes, followed by overnight incubation of ab1832 at a 1/500 dilution.
  • Overlay histogram showing Jurkat cells stained with ab1832 (blue line). The cells were prepared as follows: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight. Buffer was was 0.1% sodium azide with FBS in PBS. Cells fixed with paraformaldehyde and then permeabilized with Triton X-100 and NP40. Cell population was gated by isolating cell population from plot of SSC-A / FSA-A. The cells were then incubated the antibody at 1/100 for 24 hours. The secondary antibody used was FITC-conjugated Goat anti-Mouse polyclonal at 1/100 dilution. Isotype control antibody (red line) was used under the same conditions

    See Abreview

  • Anti-DNA PKcs antibody ab1832 used to stain Human tonsil tissue. The antibody requires antigen retrieval with citrate buffer and we recommend a 1:50-1:100 dilution with a 30 minute incubation time of the antibody at room temperature.

References for Anti-DNA PKcs antibody [18-2] (ab1832)

This product has been referenced in:
  • Zhou Y  et al. Quantitation of DNA double-strand break resection intermediates in human cells. Nucleic Acids Res 42:e19 (2014). WB ; Human . Read more (PubMed: 24362840) »
  • Sahlberg SH  et al. The influence of AKT isoforms on radiation sensitivity and DNA repair in colon cancer cell lines. Tumour Biol 35:3525-34 (2014). ICC/IF ; Human . Read more (PubMed: 24338765) »

See all 10 Publications for this product

Product Wall

Application Immunohistochemistry (Frozen sections)
Blocking step 0.25%TritonX100, 0.2%Gelatine as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
Sample Mouse Tissue sections (mouse embryonic spinal cord E11)
Specification mouse embryonic spinal cord E11
Permeabilization Yes - 0.25%TritonX100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 07 2014

DISCOUNT CODE: ***
Expiration date: November 6, 2012

This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview forab1832 in ChIPand include this code in the “Additio...

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- to optimize the transfer conditions by increasing the transfer time to 24 hours (in some examples, for big proteins, the transfer is done for 48 hours)
-a "boost" at 70-80V for 1 hour at the end of the transfer can be of sign...

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Thank you for contacting us regarding this. I've spoken with the lab regarding our Anti-DNA PKcs antibodies and have found that  ab1832 would most likely fulfill your needs as it's epitope site is between  aa1-2713. I hope this information is helpful t...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (Jurkat Cell)
Specification Jurkat Cell
Preparation Cell harvesting/tissue preparation method: Spin down, wash in FACS buffer 1x, fix, wash 2x, and stain with primary antibody overnight.
Sample buffer: Buffer composition at analysis was 0.1% sodium azide with FBS in phosphate buffered solution.
Fixation Paraformaldehyde
Permeabilization Yes - Triton X-100 and NP40
Gating Strategy Isolate cell population from plot of SSC-A / FSA-A
Username

Abcam user community

Verified customer

Submitted Sep 13 2011

Thank you for your phone call yesterday and for your patience while I looked into your question. I looked through some online resources and the best suggestion found for higher molecular weight loading controls was Hsp90 (~90 kDa). We do have some g...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Eu-Myc Arf-/- mouse lymphoma cells)
Loading amount 40 µg
Specification Eu-Myc Arf-/- mouse lymphoma cells
Treatment shRNA knockdown
Gel Running Conditions Reduced Denaturing (4%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
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Dr. Hai Jiang

Verified customer

Submitted Jun 10 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"