WB result of Phospho-DNA-PKcs (Ser2056) Recombinant Rabbit mAb
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Primary antibody incubation conditions: overnight at 4°C
Primary antibody: Phospho-DNA-PKcs (Ser2056) Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: untreated HCT 116 whole cell lysate 20 µg
Lane 2: HCT 116 treated with 0.25 µM Neocarzinoastatin for 1 hour whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 469 kDa
Observed MW: 290, 450 kDa
This blot was developed with high sensitivity substrate
Phospho-DNA-PKcs (Ser2056) Recombinant Rabbit mAb (S-3813)
Phospho-DNA-PKcs (Ser2056) Recombinant Rabbit mAb (S-3813)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Phospho-DNA-PKcs (Ser2056) |
| Synonyms | DNA-dependent protein kinase catalytic subunit; DNA-PK catalytic subunit; DNA-PKcs; DNPK1; Ser-473 kinase (S473K); p460; PRKDC; HYRC; HYRC1 |
| Location | Cytoplasm, Nucleus |
| Accession | P78527 |
| Clone Number | S-3813 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P |
| Reactivity | Hu |
| Positive Sample | HCT 116 (treated with 0.25 µM Neocarzinoastatin for 1 hour) |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu |
| IHC-P | 1:1000 | Hu |
Background
Phospho-DNA-PKcs (Ser2056) is a specific post-translational modification of the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) at serine 2056, serving as a critical hallmark of DNA-PKcs activation and the initiation of non-homologous end joining (NHEJ) repair. This phosphorylation event is primarily driven by DNA-PKcs autophosphorylation and represents an early and rapid response to DNA double-strand breaks induced by ionizing radiation or chemotherapeutic agents. Its upstream regulatory network is intricate: NEDDylation mediated by HUWE1 is a prerequisite licensing event for efficient phosphorylation at this site, whereas BRCA1, through its BRCT domain, directly binds DNA-PKcs and blocks Ser2056 autophosphorylation specifically in S-phase cells, thereby temporospatially suppressing NHEJ and channeling repair toward homologous recombination. Functionally, this modification not only drives conformational changes that promote DNA-PKcs dissociation from broken DNA ends to facilitate repair complex disassembly but also serves, in the genetic context of XLF deficiency, as a critical safeguard for end-joining fidelity during physiological V(D)J recombination in lymphocytes. Notably, Ser2056 phosphorylation exhibits remarkable non-canonical functional diversity: during mitosis, this modification occurs independently of DNA damage and the Ku heterodimer, localizing to centrosomes and the spindle apparatus to ensure proper chromosomal segregation. In the late stages of genotoxic stress, phosphorylated DNA-PKcs (Ser2056) dissociates from break sites and relocalizes to nuclear speckles, where it colocalizes with splicing factors and directly participates in the alternative splicing of pre-mRNA—establishing a direct link between DNA damage signaling and RNA metabolism. Clinically, this phospho-form is markedly upregulated and chromatin-bound in RET proto-oncogene-driven constitutively active medullary thyroid carcinoma, directly mediating chemoresistance. Meanwhile, the novel radiosensitizer black phosphorus quantum dots physically bind DNA-PKcs and specifically inhibit Ser2056 autophosphorylation, causing persistent stalling of repair complexes at damage sites and synergizing with ionizing radiation to induce apoptosis in renal cell carcinoma models.
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Western Blot
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human tonsil. Anti- Phospho-DNA-PKcs (Ser2056) antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human cervical cancer. Anti- Phospho-DNA-PKcs (Ser2056) antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti- Phospho-DNA-PKcs (Ser2056) antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human hepatocellular carcinoma. Anti- Phospho-DNA-PKcs (Ser2056) antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human gastric cancer. Anti- Phospho-DNA-PKcs (Ser2056) antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
