Anti-DNA PKcs antibody (ab70250)

Overview

  • Product nameAnti-DNA PKcs antibodySee all DNA PKcs primary antibodies ...
  • Description
    Rabbit polyclonal to DNA PKcs
  • SpecificityKu-80 co-immunoprecipitates using this antibody.
  • Tested applicationsWB, 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 a region between residue 4075 and the C-terminus (residue 4128) of human DNA PKcs. (NP_008835.5)

  • 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
IP
ICC/IF
  • Application notesIP: Use at 2 to 5 µg/mg lysate.
    WB: 1/1000 - 1/10000. Detects a band of approximately 469 kDa (predicted molecular weight: 469 kDa).


    Not yet tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 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
      • 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

    • 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

    • 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.
    • 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 for Anti-DNA PKcs antibody (ab70250)

    ab70250 has not yet been referenced specifically in any publications.

    Product Wall

    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
    Username

    Dr. Dimitra Kalamida

    Verified customer

    Submitted Dec 12 2013

    Thank you for your reply.

    I am sending you a vial of ab1832 free of charge and the new order number for this is *. Hopefully you should receive this by 10.30am tomorrow morning.

    If there is anything else I can help you with, please ...

    Read More

    Thank you for providing that information.

    I am happy to replace the DNA PKcs antibody that you currently have, free of charge. A few alternatives are listed below:

    http://www.abcam.com/DNA-PKcs-antibody-18-2-ab1832.html
    http://...

    Read More

    Thank you for your patience in this matter.

    I have still not received your emails with the IP images attached. As we discussed earlier today, it may be due to the file size being too large. Is there anyway you could reduce the size of the fil...

    Read More

    Thank you for sending this email.

    Unfortunately it does not appear that the attachments with the IP images came through. Could you please resend those images so that I may take a look at them?

    Thank you for your patience in this and...

    Read More

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