Key features and details
- Mouse monoclonal [18-2] to DNA PKcs
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Rat, Human
- Isotype: IgG1
Product nameAnti-DNA PKcs antibody [18-2]
See all DNA PKcs primary antibodies
DescriptionMouse monoclonal [18-2] to DNA PKcs
Tested applicationsSuitable for: ICC/IF, WBmore details
Species reactivityReacts with: Rat, Human
Other Immunogen Type corresponding to Human DNA PKcs aa 1-2713. Immunogen is purified from HeLa cells.
Database link: P78527
Epitopeamino acids 1-2713
- WB: Wildtype HAP1, SHSY5Y and K562 cell lysates. ICC/IF: Wildtype HAP1 cells.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.1% Sodium azide
Concentration information loading...
PurityProtein G purified
Our Abpromise guarantee covers the use of ab44815 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/200. Predicted molecular weight: 469 kDa.|
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.
- Information by UniProt
- DNA dependent protein kinase catalytic subunit antibody
- DNA PK catalytic subunit antibody
- DNA-dependent protein kinase catalytic subunit antibody
Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: DNA PKcs knockout HAP1 cell lysate (40 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: SHSY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab44815 observed at 460 kDa. Red - loading control, ab18251, observed at 52 kDa.
ab44815 was shown to specifically react with DNA PKcs in iwld-type HAP1 cells. No band was observed when DNA PKcs knockout samples were examined. Wild-type and DNA PKcs knockout samples were subjected to SDS-PAGE. Ab44815 and ab18251 (loading control to alpha tubulin) were diluted at 1/200 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
ab44815 staining DNA PKcs in wild-type HAP1 cells (top panel) and PRKDC knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab44815 at 1/100 dilution and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab44815 has been referenced in 1 publication.
- Deshpande RA et al. DNA-dependent protein kinase promotes DNA end processing by MRN and CtIP. Sci Adv 6:eaay0922 (2020). PubMed: 31934630