Product Name:
DNAPK-pY883
Product Number:
ab-pk596
Target Full Name: DNA-dependent protein kinase catalytic subunit
Target Alias: DNA-PK catalytic subunit; DNA-PKcs; DNPK1; P460; PRKD; PRKDC; XRCC7; HYRC; HYRC1; p350; ENSG00000121031
Product Type Specific: DNAPK protein kinase phosphosite-specific antibody
Antibody Code: PK596
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y883
Protein UniProt: P78527
Protein SigNET: DNAPK
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human DNAPK (PRKDC) sequence peptide Cat. No.: PE-04AFZ95
Antibody Immunogen Sequence: MKS(pY)VAW(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues M880 to W886; in the N-terminal quarter of the protein.Y883 phosphorylation stimulates phosphotransferase activity. This phosphosite is located in the kinase activation loop between catalytic subdomains VII and VIII. This is one of the major in vivo phosphorylation sites in DNAPK (≥25 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Medium immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus Macaque | Mouse | Zebra fish.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 460-480 kDa.
Related Product 1: DNAPK (880-886) pY883 antibody blocking peptide (Cat. No.: PE-04AFZ95)
Related Product 2: DNAPK S2056 phosphorylation antibody (Cat. No.: AB-PK925)
Related Product 3: DNAPK T2609 phosphorylation antibody (Cat. No.: AB-PK595)
Scientific Background: DNAPK (PRKDC) is a protein-serine/threonine kinase of the Atypical group and PIKK family. It is the 470 kDa catalytic subunit (DNA-PKcs) in a complex with a regulatory Ku70/Ku80 heterodimer that binds DNA ends, activating the kinase to protect DNA and signal repair. Phosphorylation at Thr-2609 stimulates phosphotransferase activity, whereas Ser-2056, Thr-2638, Thr-2647 and Thr-3950 phosphorylation regulates cell growth and chromatin reorganization. DNAPK has been noted to be involved in sensing DNA damage, including DNA non-homologous end joining (NHEJ) and therefore, by association, be involved with V(D)J recombination events and double-strand break (DSB) repair. It protects broken DNA ends until repair factors like Artemis and DNA ligase IV are recruited. A possible scaffolding role of DNAPK for DNA-repair enzymes has also been proposed. It participates in V(D)J recombination for creating diversity in antibodies and T-cell receptors in the immune system. A complex of Ku70 and Ku80 binds directly to broken DNA ends and recruits DNAPK to activate its kinase function. It phosphorylates targets like p53, Artemis, and itself (autophosphorylation) to regulate repair. Deficiencies in DNAPK lead to hypersensitivity to ionizing radiation and severe combined immunodeficiency (SCID) in mice. DNAPK appears to be a tumour suppressor protein (TSP). The active form of the protein kinase normally acts to inhibit tumour cell proliferation until damage to DNA can be repaired. Therefore, it can play a role to facilitate tumour cell survival under conditions of radiation and chemotherapy. Combined with cytoplasmic FUS accumulations, DNAPK is activated upon DNA damage in ALS (1). Phosphorylation of the PrLD by this protein-serine/threonine kinase inhibited the transition of FUS from a liquid to a solid phase and influenced the formation of FUS aggregates (2-4). This description may include information annotated by UniProt and/or Google AI.