Phospho-DNA-PKcs-S2056 Rabbit pAb for WB, IHC-P, IF/ICC, ELISA - P78527

Core Product Specifications and Parameters

Biological Background: DNA-PKcs Function and Localization

• DNA-PKcs (PRKDC) serves as the catalytic subunit of the DNA-dependent protein kinase complex, acting as a molecular sensor for DNA damage and playing a central role in non-homologous end joining (NHEJ) repair of double-strand breaks and V(D)J recombination. Related references: PMID:11955432 PMID:12649176 PMID:14734805
• It localizes to chromosome ends, contributing to the maintenance of telomeric stability and the prevention of chromosomal end fusion.
• Beyond DNA repair, DNA-PKcs modulates transcription, promotes ribosome biogenesis by processing precursor rRNA, and regulates innate immune responses through the cGAS-STING pathway. Related references: PMID:11955432 PMID:12649176 PMID:14734805 PMID:32103174 PMID:28712728 PMID:33273464 PMID:35460603
• It phosphorylates Ser-139 of histone H2AX, a key event in the DNA damage response, and numerous other substrates including p53, AKT, XRCC4, and WRN, thereby influencing apoptosis and cell signaling. Related references: PMID:14627815 PMID:16046194 PMID:10026262 PMID:10467406 PMID:11889123 PMID:15262962
• Predominantly a nuclear protein, DNA-PKcs is also found in the nucleolus and cytosol, reflecting its diverse functional roles.
• Subject to phosphorylation, acetylation, and ubiquitination; mutations in PRKDC are associated with severe combined immunodeficiency (SCID).

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphorylated peptide spanning Ser2056 of human DNA-PKcs, designed to ensure phosphorylation-state specificity. Include appropriate phospho-negative controls during validation.
• For Western blot, due to the very high molecular weight of DNA-PKcs (469 kDa), consider using low-percentage gels (e.g., 4–8%) and extended transfer times. Use pre-stained high-molecular-weight markers for clear visualization.
• In immunofluorescence or immunohistochemistry, anticipate strong nuclear staining along with possible nucleolar and cytoplasmic signals. Optimize fixation and permeabilization protocols to preserve the phospho-epitope.
• Cross-reactivity is reported for human; if testing other species, confirm sequence homology around Ser2056 and perform a pilot experiment.

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