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Anti-DNA PKcs (phospho T2609) antibody [10B1] (ab18356)

Overview

  • Product nameAnti-DNA PKcs (phospho T2609) antibody [10B1]See all DNA PKcs primary antibodies ...
  • Description
    Mouse monoclonal [10B1] to DNA PKcs (phospho T2609)
  • Tested applicationsICC/IF, WB more details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic modified peptide.

  • Positive controlMOLT4 nuclear extract treated with 2 to 10 Gy radiation.

Properties

Applications

Our Abpromise guarantee covers the use of ab18356 in the following tested applications.

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

Application Notes
ICC/IF ICC/IF: Use at an assay dependent concentration.
WB WB: Use a concentration of 1 µg/ml. Detects a band of approximately >400 kDa (predicted molecular weight: 460 kDa). This antibody has been seen to cross-react with a protein having an approximate molecular weight of p53. However, it is not known exactly what this protein is.

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 antibodyDNA PK catalytic subunit antibodyDNA-dependent protein kinase catalytic subunit antibody
      DNA-PK catalytic subunit antibodyDNA-PKcs antibodyDNAPK antibodyDNAPK catalytic subunit antibodyDNPK 1 antibodyDNPK1 antibodyDNPK1 antibodyHyper radiosensitivity of murine scid mutation, complementing 1 antibodyHYRC 1 antibodyHYRC antibodyHYRC1 antibodyHYRC1 antibodyp350 antibodyp460 antibodyPKRDC antibodyPRKDC antibodyPRKDC_HUMAN antibodyProtein Kinase DNA Activated Catalytic Polypeptide antibodyProtein Kinase DNA Activated Catalytic Polypeptide antibodyXRCC 7 antibodyXRCC7 antibodyXRCC7 antibody
    see all

Anti-DNA PKcs (phospho T2609) antibody [10B1] images



  • Predicted band size : 460 kDa


    MOLT4 nuclear extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with anti-DNAPKcs (Thr2609) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Lane 1, untreated MOLT4 cells. Lane 2, MOLT4 cells exposed to 2 Gy radiation. Lane 3, MOLT4 cells exposed to 10 Gy radiation. In both cases, cells were harvested 30 minutes after radiation.

References for Anti-DNA PKcs (phospho T2609) antibody [10B1] (ab18356)

This product has been referenced in:
  • Yue J  et al. Identification of the DNA repair defects in a case of Dubowitz syndrome. PLoS One 8:e54389 (2013). ICC/IF ; Human . Read more (PubMed: 23372718) »
  • Solier S  et al. Heat shock protein 90a (HSP90a), a substrate and chaperone of DNA-PK necessary for the apoptotic response. Proc Natl Acad Sci U S A 109:12866-72 (2012). ICC/IF . Read more (PubMed: 22753480) »

See all 11 Publications for this product

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Thank you for confirming these details and for your cooperation.
As requested, I have issued a free of charge replacement of 2 units of ab18356 with ab97611 and ab18192.
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Thank you very much for your interest in ab18356, ab4194, and ab97611.

To our knowledge, ab18356, ab4194, and ab97611 have not been tested in mouse. We don't have smaller sizes of our antibodies, but I can offer a discount off a future purchase...

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Thank you for those precisions.
Yes, I think a transfer overnight at 30V followed by a "boost" at 70-80V for 1 hour at the end of the transfer can be of significant improvement. I would recommend to add 0.1% SDS in the transfer buffer.
A gradien...

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Thank you for taking time to complete our questionnaire. I am sorry to hear that you are experiencing some problems with this antibody in Western Blot.
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Vielen Dank für diese Rückmeldung! Wir würden uns über ein Feedback von Ihnen, wie die Versuche mit ab18356 geklappt haben, sehr freuen: In Ihrem Fall könnte ich nämlich eine Ausnahme machen und Ihnen den Rabattcode auch noch nachträglich ausstellen. Sie ...

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Vielen Dank für Ihren Anruf und für Ihr Interesse an unseren Produkten. Leider konnte ich Sie in den letzten Tagen telefonisch nicht erreichen. Daher hoffe ich, mit dieser email Ihre Frage beantworten zu können: Unseres Wissens nach wurde der anti-D...

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Application Western blot
Sample Human Cell lysate - whole cell (HeLa and mouse embrionic fibroblasts (MEFs))
Loading amount 40 µg
Specification HeLa and mouse embrionic fibroblasts (MEFs)
Treatment 10Gy for HeLa 30Gy for MEFs
Gel Running Conditions Reduced Denaturing (6% SDS-Page)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Apr 12 2010



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