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AB195537

HRP Anti-DNA PKcs antibody [Y393]

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Rabbit Recombinant Monoclonal DNA PKcs antibody - conjugated to HRP. Suitable for WB and reacts with Human samples.

View Alternative Names

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

2 Images
Western blot - HRP Anti-DNA PKcs antibody [Y393] (AB195537)
  • WB

Lab

Western blot - HRP Anti-DNA PKcs antibody [Y393] (AB195537)

ab195537 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in PRKDC knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PRKDC knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab195537 and ab7291 (Mouse anti Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

All lanes:

Western blot - HRP Anti-DNA PKcs antibody [Y393] (ab195537) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 40 µg

Lane 2:

PRKDC knockout HAP1 whole cell lysate at 40 µg

Predicted band size: 469 kDa

Observed band size: 460 kDa

false

Western blot - HRP Anti-DNA PKcs antibody [Y393] (AB195537)
  • WB

Lab

Western blot - HRP Anti-DNA PKcs antibody [Y393] (AB195537)

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab195537 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

All lanes:

Western blot - HRP Anti-DNA PKcs antibody [Y393] (ab195537) at 1/5000 dilution

All lanes:

K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 10 µg

Predicted band size: 469 kDa

Observed band size: 460 kDa

true

Exposure time: 20min

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

Y393

Isotype

IgG

Conjugation

HRP

Excitation/Emission
Carrier free

No

Reacts with

Human

Applications

WB

applications

Immunogen

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

Reactivity data

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Product details

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.4 Preservative: 0.1% Proclin 300 Solution Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
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|Store in the dark

Supplementary information

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.

Product protocols

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

Target data

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

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