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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.


Images

Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (AB18192), expandable thumbnail
  • Western blot - Anti-DNA PKcs (phospho S2056) antibody (AB18192), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (AB18192), expandable thumbnail
  • ELISA - Anti-DNA PKcs (phospho S2056) antibody (AB18192), expandable thumbnail
  • Western blot - Anti-DNA PKcs (phospho S2056) antibody (AB18192), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA

Form

Liquid

Clonality

Polyclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Consider this alternative

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ELISAWBICC/IF
Human
Expected
Tested
Expected
Mouse
Predicted
Predicted
Predicted
Synthetic peptide - Human
Tested
Not recommended
Not recommended

Tested
Tested

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).

Expected
Expected

Species

Human

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Mouse

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1 µg/mL

Notes

-

Predicted
Predicted

Species

Mouse

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Expected
Expected

Species

Human

Dilution info

1-5 µg/mL

Notes

-

Predicted
Predicted

Species

Mouse

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Synthetic peptide - Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

8 products for Alternative Product

Target data

Function

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).

Alternative names

Recommended products

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.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Polyclonal

Immunogens
  • The exact immunogen used to generate this antibody is proprietary information.
Purification technique

Affinity purification Immunogen

Specificity

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.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Abcam is leading the way to address reproducibility in scientific research with our highly validated recombinant monoclonal and recombinant multiclonal antibodies. Search & select one of Abcam's thousands of recombinant alternatives to eliminate batch-variability and unnecessary animal use.

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Abcam antibodies are extensively validated in a wide range of species and applications, so please check the reagent specifications meet your scientific needs before purchasing. If you have any questions or bespoke requirements, simply visit the Contact Us page to send us an inquiry or contact our Support Team ahead of purchase.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

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6 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail
    Furusawa Y et al. DNA double-strand breaks induced by cavitational mechanical effects of ultrasound in cancer cell lines. PLoS One 7:e29012 (2012). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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.

  • Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail

    Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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.

  • ELISA - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail

    ELISA - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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.

  • Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail

    Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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

    Secondary

    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

  • Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192), expandable thumbnail

    Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192)

    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

    Secondary

    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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