Anti-DNA PKcs antibody [18-2] (ab1832)
- Product nameAnti-DNA PKcs antibody [18-2]See all DNA PKcs primary antibodies ...
- DescriptionMouse 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 Cyt more details
- Species reactivityReacts with: Mouse, Rat, Human
Human DNA-dependent Protein Kinase purified from HeLa cells.
- EpitopeAmino acids 1-2713.
- Positive controlLS174T cells. Tonsil.
- Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
- Storage buffer10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
- Concentration information loading...
- PurityProtein G purified
- Clonality Monoclonal
- Clone number18-2
- Research Areas
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.
|Functional Studies||FuncS: Use at an assay dependent dilution.|
|IHC-P||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||IHC-Fr: Use at an assay dependent dilution.|
|IP||IP: Use at an assay dependent dilution. (protein G) Use at a concentration of 2µg/mg of protein lysate (native and denatured).|
|WB||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||ICC/IF: Use at an assay dependent dilution. PubMed: 20709753|
|Flow Cyt||Flow Cyt: Use at an assay dependent concentration.|
- 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.
modificationsPhosphorylated 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.
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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
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:
- Sanz-García M et al. Vaccinia-related Kinase 1 (VRK1) Is an Upstream Nucleosomal Kinase Required for the Assembly of 53BP1 Foci in Response to Ionizing Radiation-induced DNA Damage. J Biol Chem 287:23757-68 (2012). Read more (PubMed: 22621922) »
- Liccardi G et al. EGFR nuclear translocation modulates DNA repair following cisplatin and ionizing radiation treatment. Cancer Res 71:1103-14 (2011). WB, ICC/IF ; Mouse . Read more (PubMed: 21266349) »