AB18192

Anti-DNA PKcs (phospho S2056) 抗体

Name: Anti-DNA PKcs (phospho S2056) 抗体 (ab18192) | Abcam
Brand: Abcam Limited
SKU: AB18192
Price: 59000 JPY
Availability: InStock
Rating: 4 (14 reviews)

Anti-DNA PKcs (phospho S2056) antibody

KO Validated
詳細を見る

(14 Reviews)

(229 Publications)

Anti-DNA PKcs (phospho S2056) antibody (ab18192) is a rabbit polyclonal antibody detecting DNA PKcs in Western Blot, ICC/IF, ELISA. Suitable for Human.

- KO validated for confirmed specificity
- Over 190 publications
- Trusted since 2006

別名を表示する

HYRC, HYRC1, PRKDC, DNA-dependent protein kinase catalytic subunit, DNA-PK catalytic subunit, DNA-PKcs, DNPK1, Ser-473 kinase, p460, S473K

7 Images

Lab

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, 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (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)

Predicted band size: 469 kDa

Observed band size: 117 kDa,150 kDa,450 kDa,85 kDa

false

Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (AB18192)

ICC/IF

Lab

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 ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and 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.

ICC/IF

PubMed

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.

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/

Unknown

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, 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

Lane 1:

Gamma Irradiated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Gamma Irradiated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406)

Lane 4:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406)

Lane 5:

Gamma Irradiated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with Human DNA PKcs peptide (<a href='/products/proteins-peptides/human-dna-pkcs-peptide-ab20407'>ab20407</a>)

Lane 6:

Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with Human DNA PKcs peptide (<a href='/products/proteins-peptides/human-dna-pkcs-peptide-ab20407'>ab20407</a>)

Secondary

All lanes:

Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution

Predicted band size: 469 kDa

Observed band size: 270 kDa,460 kDa

true

ELISA - Anti-DNA PKcs (phospho S2056) antibody (AB18192)

ELISA

Lab

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 (ab97080; diluted 50000 times) and signal was developed with TMB substrate.

Project

Western blot - Anti-DNA PKcs (phospho S2056) antibody (AB18192)

All lanes:

Western blot - Anti-DNA PKcs (phospho S2056) antibody (ab18192) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) at 20 µg

Lane 3:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406)

Lane 4:

Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) at 20 µg with Human DNA PKcs (phospho S2056) peptide (ab20406)

Lane 5:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg with Human DNA PKcs peptide (<a href='/products/proteins-peptides/human-dna-pkcs-peptide-ab20407'>ab20407</a>)

Lane 6:

Hela Whole Cell Lysate - Bleomycin Treated (20U/ml) at 20 µg with Human DNA PKcs peptide (<a href='/products/proteins-peptides/human-dna-pkcs-peptide-ab20407'>ab20407</a>)

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 469 kDa

Observed band size: 117 kDa,150 kDa,270 kDa,460 kDa,50 kDa

false

Exposure time: 15min

ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs (phospho S2056) antibody (AB18192)

Immunocytochemistry-immunofluorescence using Anti-DNA PKcs (phospho S2056) antibody, ab18192. Publication image from Gustafsson, N. M. S. et al., 2018, Nat Commun, 30250201. Legend direct from paper.

ATM signaling stimulates PFKFB3 recruitment upon DNA damage induction. a Confocal analysis of DNA damage, γH2AX, and nuclear localization of PFKFB3 in U2OS cells following treatment with inhibitors (6 h) as indicated, subjected to IR (6 Gy), or left untreated, n = 3 independent experiments. Scale bar, 10 µm. b Scatter dot plot representing the PFKFB3 nuclear intensity in (a) as quantified using CellProfiler, n > 100 cells/treatment. ***P < 0.001; one-way ANOVA analysis. c Scatter dot plot representing the γH2AX nuclear intensity in (a) as quantified using CellProfiler, n > 100 cells/treatment. **P < 0.01, ***P < 0.001; one-way ANOVA analysis. d Bars representing the percentage of PFKFB3 and γH2AX foci that co-localize in (a) as quantified using CellProfiler, n > 100 cells/treatment. Data are displayed as means ± SEM, ***P < 0.001, one-way ANOVA was used to calculate statistical significance

Key facts

宿主種

Rabbit

クローン性

Polyclonal

アイソタイプ

IgG

キャリアフリー

交差種

Human

アプリケーション

WB, ICC/IF, ELISA

applications

免疫原

This product was produced with the following immunogens:

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

特異性

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.

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Reactivity data

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製品の詳細

What is this antibody validated in?
Anti-DNA PKcs (phospho S2056) antibody (ab18192) is a rabbit polyclonal antibody and is validated for use in Western Blot (WB), Immunocytochemistry/immunofluorescence (ICC/IF), ELISA in Human samples.

What is the molecular weight of DNA PKcs?
Anti-DNA PKcs (phospho S2056) (ab18192) specifically detects a band for DNA PKcs (UniProt: P78527) at a molecular weight of 460kDa.

Trusted by the scientific community
Anti-DNA PKcs (phospho S2056) (ab18192) was first used in a scientific publication in 2006 and has been cited over 190 times in peer-reviewed journals.

Reviewed by scientists
Anti-DNA PKcs (phospho S2056) (ab18192) has over 10 independent reviews from customers.

Specificity confirmed
The specificity of Anti-DNA PKcs (phospho S2056) antibody (ab18192) has been confirmed by Western blot testing in DNA PKcs Knockout HAP1 cells.

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出荷温度及び保存条件

製品の状態

Liquid

精製方法

Affinity purification Immunogen

バッファー組成

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

出荷温度

Blue Ice

短期保存期間

1-2 weeks

短期保存温度

+4°C

長期保存温度

-20°C

分注に関する情報

Upon delivery aliquot

保管に関する情報

Avoid freeze / thaw cycle

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補足情報

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.

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.

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.

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製品プロトコール

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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ターゲットの情報

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).

See full target information PRKDC phospho S2056

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文献 (229)

Recent publications for all applications. Explore the full list and refine your search

Nucleic acids research 53: PubMed40548939

2025

USP37 counteracts HLTF to protect damaged replication forks and promote survival of BRCA1-deficient cells and PARP inhibitor resistance.

Applications

Unspecified application

Species

Unspecified reactive species

Mengfan Tang,Siting Li,Ziqiang Zhu,Chao Wang,Shuai Ding,Huimin Zhang,Min Huang,Sarah Keast,Zhen Chen,Ling Yin,Litong Nie,Dan Su,Xu Feng,Junjie Chen

Molecular medicine (Cambridge, Mass.) 31:222 PubMed40484931

2025

Nuclear VPS35 attenuates NHEJ repair by sequestering Ku protein.

Applications

Unspecified application

Species

Unspecified reactive species

Luping Zhang,Yonghong Nie,Tuo Tang,Yanji Lu,Wenlong Li,Xian Hong,Qiang Li,Aixue Zheng,Yongpei Li,Jianwen Zhou,Li Fan,Tao Wang,Zhihui Deng

JCI insight 10: PubMed40392611

2025

Loss of genome maintenance is linked to mTOR complex 1 signaling and accelerates podocyte damage.

Applications

Unspecified application

Species

Unspecified reactive species

Fabian Braun,Amrei M Mandel,Linda Blomberg,Milagros N Wong,Georgia Chatzinikolaou,David H Meyer,Anna Reinelt,Viji Nair,Roman Akbar-Haase,Phillip J McCown,Fabian Haas,He Chen,Mahdieh Rahmatollahi,Damian Fermin,Robin Ebbestad,Gisela G Slaats,Tillmann Bork,Christoph Schell,Sybille Koehler,Paul T Brinkkoetter,Maja T Lindenmeyer,Clemens D Cohen,Martin Kann,David Unnersjö-Jess,Wilhelm Bloch,Matthew G Sampson,Martijn Et Dollé,Victor G Puelles,Matthias Kretzler,George A Garinis,Tobias B Huber,Bernhard Schermer,Thomas Benzing,Björn Schumacher,Christine E Kurschat

Aging cell 24:e70079 PubMed40371563

2025

The Redox Activity of Protein Disulphide Isomerase Functions in Non-Homologous End-Joining Repair to Prevent DNA Damage.

Applications

Unspecified application

Species

Unspecified reactive species

Sina Shadfar,Fabiha Farzana,Sayanthooran Saravanabavan,Ashley M Rozario,Marta Vidal,Cyril Jones Jagaraj,Sonam Parakh,Esmeralda Paric,Kristy C Yuan,Mariana Brocardo,Donna R Whelan,Angela S Laird,Julie D Atkin

Science advances 11:eado7660 PubMed40238889

2025

Nuclear accumulation of YTHDF1 regulates mRNA splicing in the DNA damage response.

Applications

Unspecified application

Species

Unspecified reactive species

Jingyu Hou,Yunyi Gao,Bing Han,Sujun Yan,Saisai Wei,Xiangwei Gao

Cell death discovery 11:133 PubMed40175326

2025

Dual role of p21 in regulating apoptosis and mitotic integrity in response to doxorubicin in colon cancer cells.

Applications

Unspecified application

Species

Unspecified reactive species

Heeyeon Kim,Haein Kim,Eunjung Jang,Young-Woo Eom,Gyesoon Yoon,Kyeong Sook Choi,Eunhee Kim

Cell death & disease 16:49 PubMed39870644

2025

Absent in melanoma 2: a potent suppressor of retinal pigment epithelial-mesenchymal transition and experimental proliferative vitreoretinopathy.

Applications

Unspecified application

Species

Unspecified reactive species

Yu Chen,Mingyuan Jiang,Liping Li,Shanshan Yang,Zuimeng Liu,Shiwen Lin,Wanxiao Wang,Jinyang Li,Feng Chen,Qiang Hou,Xiaoyin Ma,Ling Hou

Nature cancer 6:278-291 PubMed39838098

2025

Targeting BRCA1-deficient PARP inhibitor-resistant cells with nickases reveals nick resection as a cancer vulnerability.

Applications

Unspecified application

Species

Unspecified reactive species

Jenna M Whalen,Jillian Earley,Christi Silva,Arthur M Mercurio,Sharon B Cantor

Science advances 11:eads6215 PubMed39772686

2025

Infection-induced lysine lactylation enables herpesvirus immune evasion.

Applications

Unspecified application

Species

Unspecified reactive species

Matthew D Tyl,Victoria U Merengwa,Ileana M Cristea

Translational oncology 52:102231 PubMed39662449

2024

CDK4/6 inhibitors promote PARP1 degradation and synergize with PARP inhibitors in non-small cell lung cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Carlos M Roggero,Anwesha B Ghosh,Anvita Devineni,Shihong Ma,Eliot Blatt,Ganesh V Raj,Yi Yin

View all publications

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