Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)
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Overview
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Product nameAnti-DNA PKcs antibody [EPR392] - BSA and Azide free
See all DNA PKcs primary antibodies -
DescriptionRabbit monoclonal [EPR392] to DNA PKcs - BSA and Azide free
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC/IFmore details
Unsuitable for: Flow Cyt,IHC-P or IP -
Species reactivityReacts with: Human
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Positive control
- WB: K562, Molt4, MCF7, SH-SY5Y, 293T and PC3 cell lysates. ICC/IF: K-562 cells
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General notes
The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).
ab174573 is a PBS-only buffer formulated version of ab133516, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab133516 for information on protocols, dilutions, and image data.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
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.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
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Storage bufferConstituent: PBS
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Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR392
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IsotypeIgG
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Research areas
Associated products
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Alternative Products
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Alternative Versions
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab174573 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 | Use at an assay dependent concentration. Detects a band of approximately 460 kDa (predicted molecular weight: 469 kDa). | |
| ICC/IF | Use at an assay dependent concentration. |
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
- Omim: 600899 Human
- SwissProt: P78527 Human
- Unigene: 491682 Human
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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
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Images
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![Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.com/ps/products/174/ab174573/Images/ab174573-309380-ab133516KOWB.jpg)
Western blot - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)This WB data was generated using the same anti-DNA PKcs antibody clone, EPR392, in a different buffer formulation (cat# ab133516).
Lane 1: Wild type HAP1 whole cell lysate (40 µg)
Lane 2: DNA PKcs knockout HAP1 whole cell lysate (40 µg)
Lane 3: K562 whole cell lysate (20 µg)
Lane 4: SHSY5Y whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab133516 observed at 460 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab133516 was shown to specifically react with DNA PKcs when DNA PKcs knockout samples were used. Wild-type and DNA PKcs knockout samples were subjected to SDS-PAGE. Ab133516 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.com/ps/products/174/ab174573/Images/ab174573-3-ab133516ICCIF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)Immunocytochemistry/ Immunofluorescence analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labeling DNA PKcs with purified ab133516 at 1:200 dilution (9.3 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
![Other - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)](https://www.abcam.com/ps/products/174/ab174573/Images/ab174573-309379-ab133516Affinity.jpg)
Other - Anti-DNA PKcs antibody [EPR392] - BSA and Azide free (ab174573)This kD data was generated using the same anti-DNA PKcs antibody clone, EPR392, in a different buffer formulation (cat# ab133516).
Equilibrium disassociation constant (KD)
Learn more about KD
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Datasheets and documents
References
ab174573 has not yet been referenced specifically in any publications.
