Anti-DNA PKcs antibody [Y393] (ab32566)
- Datasheet
- Specific References (11)
- Protocols
Overview
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Product nameAnti-DNA PKcs antibody [Y393]
See all DNA PKcs primary antibodies -
DescriptionRabbit monoclonal [Y393] to DNA PKcs
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Host speciesRabbit
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SpecificityThis antibody is specific for DNA PKcs. It may also detect the splice isoform 2.
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Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP -
Species reactivityReacts with: Mouse, Rat, Human, Armenian hamster
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Immunogen
Synthetic peptide within Human DNA PKcs aa 4050 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P78527 -
Positive control
- WB: K562 cell lysate. ICC/IF: Hela cells. IHC-P: Human breast carcinoma tissue slides.
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General notes
Mouse and rat samples are recommended based on WB results.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberY393
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab32566 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/1000 - 1/10000. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa). For unpurified, use 1/1000 - 1/2000. |
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| IHC-P | 1/50. For unpurified, use 1/5. |
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| ICC/IF | 1/100. For unpurified, use 1/10. |
Target
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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.
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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. -
Post-translational
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.
- Information by UniProt
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Database links
- Entrez Gene: 5591 Human
- Entrez Gene: 19090 Mouse
- Entrez Gene: 360748 Rat
- Omim: 600899 Human
- SwissProt: P78527 Human
- SwissProt: P97313 Mouse
- Unigene: 491682 Human
- Unigene: 71 Mouse
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Alternative names
- DNA dependent protein kinase catalytic subunit antibody
- DNA PK catalytic subunit antibody
- DNA-dependent protein kinase catalytic subunit antibody
see all
Images
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![Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-266738-ab32566KOWB.jpg)
Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: DNA PKcs knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lane 4: SH-SY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32566 observed at 470 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab32566 was shown to specifically react with DNA PKCs in wild-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. ab32566 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and 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. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239368-32566IHC2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)Immunohistochemical staining of paraffin embedded human tonsil with purified ab32566 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
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![Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239373-32566-ICC-2.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [Y393] (ab32566)Immunofluorescent staining of HeLa cells (fixed with 4% PFA) with purified ab32566 at a dilution of 1/100. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.
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![Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239366-32566-WB-2.jpg)
Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)All lanes : Anti-DNA PKcs antibody [Y393] (ab32566) at 1/7300 dilution (purified)
Lane 1 : MOLT4 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 469 kDa
Observed band size: 460 kDa (why is the actual band size different from the predicted?)Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239367-32566IHC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)Immunohistochemical staining of paraffin embedded human tonsil with unpurified ab32566 at a working dilution of 1 in 5. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
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![Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239371-32566-ICC-1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [Y393] (ab32566)Immunofluorescent staining of HeLa cells (fixed with 4% PFA) with unpurified ab32566 at a dilution of 1/10. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.
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![Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566-239364-32566-WB-1.jpg)
Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)All lanes : Anti-DNA PKcs antibody [Y393] (ab32566) at 1/1000 dilution (unpurified)
Lane 1 : MOLT4 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 469 kDa
Observed band size: 460 kDa (why is the actual band size different from the predicted?)Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566_2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNA PKcs antibody [Y393] (ab32566)Unpurified ab32566, at a 1/50 dilution, staining DNA PKcs in paraffin embedded human breast carcinoma tissue by immunohistochemistry.
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![Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)](https://www.abcam.com/ps/products/32/ab32566/Images/ab32566_1.jpg)
Western blot - Anti-DNA PKcs antibody [Y393] (ab32566)Anti-DNA PKcs antibody [Y393] (ab32566) at 1/1000 dilution (unpurified) + K562 cell lysate.
Predicted band size: 469 kDa
Observed band size: 460 kDa (why is the actual band size different from the predicted?)
References
This product has been referenced in:
- Sun JS & Yang XH Expression of DNA-dependent protein kinase catalytic subunit in laryngeal squamous cell carcinoma and its importance. Exp Ther Med 15:3295-3301 (2018). Read more (PubMed: 29545847) »
- Sekiba K et al. DHX9 regulates production of hepatitis B virus-derived circular RNA and viral protein levels. Oncotarget 9:20953-20964 (2018). Read more (PubMed: 29765512) »
Customer reviews and Q&As
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