Knockout Tested Rabbit Polyclonal DNA PKcs phospho S2056 antibody. Suitable for ELISA, WB, ICC/IF and reacts with Synthetic peptide - Human, Human samples. Cited in 195 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
ELISA | WB | ICC/IF | |
---|---|---|---|
Human | Expected | Tested | Expected |
Mouse | Predicted | Predicted | Predicted |
Synthetic peptide - Human | Tested | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes Direct ELISA with serially diluted ab18192 (0.2-1000 ng x mL-1), bound to immobilised phospho or control peptides (1 μg x mL-1). |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1-5 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Synthetic peptide - Human | Dilution info - | Notes - |
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Serine/threonine-protein kinase that acts as a molecular sensor for DNA damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234). Involved in DNA non-homologous end joining (NHEJ) required for double-strand break (DSB) repair and V(D)J recombination (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:33854234, PubMed:34352203). Must be bound to DNA to express its catalytic properties (PubMed:11955432). Promotes processing of hairpin DNA structures in V(D)J recombination by activation of the hairpin endonuclease artemis (DCLRE1C) (PubMed:11955432). Recruited by XRCC5 and XRCC6 to DNA ends and is required to (1) protect and align broken ends of DNA, thereby preventing their degradation, (2) and sequester the DSB for repair by NHEJ (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326, PubMed:33854234). Act as a scaffold protein to aid the localization of DNA repair proteins to the site of damage (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). The assembly of the DNA-PK complex at DNA ends is also required for the NHEJ ligation step (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Found at the ends of chromosomes, suggesting a further role in the maintenance of telomeric stability and the prevention of chromosomal end fusion (By similarity). Also involved in modulation of transcription (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). As part of the DNA-PK complex, involved in the early steps of ribosome assembly by promoting the processing of precursor rRNA into mature 18S rRNA in the small-subunit processome (PubMed:32103174). Binding to U3 small nucleolar RNA, recruits PRKDC and XRCC5/Ku86 to the small-subunit processome (PubMed:32103174). Recognizes the substrate consensus sequence [ST]-Q (PubMed:11955432, PubMed:12649176, PubMed:14734805, PubMed:15574326). Phosphorylates 'Ser-139' of histone variant H2AX, thereby regulating DNA damage response mechanism (PubMed:14627815, PubMed:16046194). Phosphorylates ASF1A, DCLRE1C, c-Abl/ABL1, histone H1, HSPCA, c-jun/JUN, p53/TP53, PARP1, POU2F1, DHX9, FH, SRF, NHEJ1/XLF, XRCC1, XRCC4, XRCC5, XRCC6, WRN, MYC and RFA2 (PubMed:10026262, PubMed:10467406, PubMed:11889123, PubMed:12509254, PubMed:14599745, PubMed:14612514, PubMed:14704337, PubMed:15177042, PubMed:1597196, PubMed:16397295, PubMed:18644470, PubMed:2247066, PubMed:2507541, PubMed:26237645, PubMed:26666690, PubMed:28712728, PubMed:29478807, PubMed:30247612, PubMed:8407951, PubMed:8464713, PubMed:9139719, PubMed:9362500). Can phosphorylate C1D not only in the presence of linear DNA but also in the presence of supercoiled DNA (PubMed:9679063). Ability to phosphorylate p53/TP53 in the presence of supercoiled DNA is dependent on C1D (PubMed:9363941). Contributes to the determination of the circadian period length by antagonizing phosphorylation of CRY1 'Ser-588' and increasing CRY1 protein stability, most likely through an indirect mechanism (By similarity). Plays a role in the regulation of DNA virus-mediated innate immune response by assembling into the HDP-RNP complex, a complex that serves as a platform for IRF3 phosphorylation and subsequent innate immune response activation through the cGAS-STING pathway (PubMed:28712728). Also regulates the cGAS-STING pathway by catalyzing phosphorylation of CGAS, thereby impairing CGAS oligomerization and activation (PubMed:33273464). Also regulates the cGAS-STING pathway by mediating phosphorylation of PARP1 (PubMed:35460603).
HYRC, HYRC1, HYRC1, PRKDC, HYRC, DNA-dependent protein kinase catalytic subunit, DNA-PK catalytic subunit, DNA-PKcs, DNPK1, p460
Knockout Tested Rabbit Polyclonal DNA PKcs phospho S2056 antibody. Suitable for ELISA, WB, ICC/IF and reacts with Synthetic peptide - Human, Human samples. Cited in 195 publications.
IgG
Rabbit
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
Liquid
Polyclonal
Affinity purification Immunogen
From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. This antibody specifically recognizes a band at ~460 kDa in HeLa cells that have been treated with ionizing radiation, that is not detected in untreated cells. This band can also be competed away by the immunizing modified peptide, but not the unmodified peptide containing the same amino acid sequence.All batches of this antibody are screened in ELISA and show high titres against the immunising peptide. Reactivity with the unmodified DNA PKcs peptide is minimal.We now predict that this antibody will cross react with mouse DNA PKcs, as the mouse sequence has been more extensively reviewed on uniprot (P97313), now indicating that mouse DNA PKcs S2053, which corresponds to human S2056, is also phosphorylated. We welcome any feedback from researchers who have used this antibody with mouse samples.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
DNA-dependent protein kinase catalytic subunit often abbreviated as DNA-PKcs or PKcs is an important component in the cellular machinery responsible for DNA repair. This protein weighs approximately 470 kDa and is predominantly expressed in various tissue types with higher levels found in lymphoid tissues. The DNA-PKcs functions as a serine/threonine protein kinase and becomes active upon binding to DNA contributing to its role in maintaining genome integrity.
DNA-PKcs plays an essential role in the non-homologous end joining (NHEJ) repair pathway which repairs double-strand breaks in DNA. DNA-PKcs forms a complex with the Ku70/80 heterodimer serving as a critical component of the DNA repair mechanism. This complex recognizes DNA ends facilitates their synapsis and activates other enzymes involved in ligating the broken DNA strands therefore promoting cellular survival following DNA damage.
DNA-PKcs deeply integrates into the DNA damage response pathway. It collaborates closely with proteins such as ATM (ataxia-telangiectasia mutated) to sense DNA damage and initiate repair. DNA-PKcs is involved in V(D)J recombination vital for the generation of diverse antibodies in immune cells. Its activity is essential for coordinating with other proteins including XRCC4 and Ligase IV to ensure efficient DNA repair processes are executed.
Defects in DNA-PKcs are linked to severe combined immunodeficiency (SCID) due to its central role in V(D)J recombination. Dysfunction in its pathway also relates to cancer where impaired DNA repair mechanisms substantially increase genomic instability leading to tumorigenesis. DNA-PKcs interacts with other proteins like BRCA1 and p53 in tumors where its overexpression or mutations can contribute to resistance against radiation and chemotherapeutic agents. These interactions highlight the importance of DNA-PK inhibitors as potential therapeutic strategies in oncology.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Full details and terms and conditions can be found here:
Terms & Conditions.
U937 (Human histiocytic lymphoma cell line) cells were paraformaldehyde-fixed. Control and treated cells were permeabilized/blocked with 2% BSA/0.05% Triton X-100/Tris-buffered saline, and immunostained for DNA PKcs (phospho S2056) (Red) for 2 h with ab18192 at 1/600 dilution.
Green staining shows γH2AX. IR = Ionzing radiation. US = Ultrasound.
Lane 1: Wild type HAP1 whole cell lysate (20 μg)
Lane 2: HAP1 Parental Camptothecin (1um 1hr) whole cell lysate (20 μg)
Lane 3: HAP1 PRKDC KO Untreated whole cell lysate (20 μg)
Lane 4: HAP1 PRKDC KO camptothecin (1um 1hr) whole cell lysate (20ug)
Lane 5: SHSY-5Y untreated whole cell lysate (20ug)
Lane 6: SHSY-5Y Camptothecin treated (1uM, 1hr) whole tissue lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab18192 observed at 450 kDa. Red - loading control, Anti-beta Catenin antibody [15B8] ab6301, observed at 85 kDa.
ab18192 was shown to specifically react with DNA PKcs (phospho S2056) in wild-type HAP1 cells along with additional cross reactive bands. Uplift was also seen with Camptothecin treatment and no bands were observed when DNA PKcs (phospho S2056) knockout cellsd. Wild-type and DNA PKcs (phospho S2056) knockout samples were subjected to SDS-PAGE. ab18192 and Anti-beta Catenin antibody [15B8] ab6301 (Mouse anti-Beta Catenin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192)
Performed under reducing conditions.
Predicted band size: 469 kDa
Observed band size: 117 kDa, 150 kDa, 450 kDa, 85 kDa
ab18192 staining DNA PKcs (phospho S2056) in HeLa UV cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 overnight at 4°C with ab18192 at 1μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Serially diluted ab18192 was bound to immobilised DNA PKcs phospho peptide (2052 - 2062; P-S2056) or DNA PKcs control peptide (2052 - 2062; both at 1 microgram x mL-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080; diluted 50000 times) and signal was developed with TMB substrate.
All lanes: Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192) at 1 µg/mL
Lanes 1, 3 and 5: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg
Lanes 2, 4 and 6: Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) at 20 µg
All lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 469 kDa
Observed band size: 117 kDa, 150 kDa, 270 kDa, 460 kDa, 50 kDa
Exposure time: 15min
ab18192 specifically recognizes a band at ~460 kDa corresponding to DNA PKcs in HeLa cells that have been treated with ionizing radiation (lane 1). This band is not detected in untreated cells (lane 2). The activity of the antibody is quenched by the addition of the immunizing (modified) peptide, ab20406 (lanes 3) but not the unmodified peptide, Human DNA PKcs peptide ab20407 (lane 5).
For the ab13823 irradiated HeLa cell lysate, the 4 hour post-treatment extract was used.
All lanes: Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192) at 1 µg/mL
Lanes 1, 3 and 5: Gamma Irradiated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lanes 2, 4 and 6: Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
All lanes: Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 469 kDa
Observed band size: 270 kDa, 460 kDa
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