PRKDC Antibody

Rabbit anti-DNA PKcs pT2609 Antibody, DNA dependent protein kinase catalytic subunit antibody, Protein Kinase DNA-Activated Catalytic Polypeptide, DNA PK catalytic subunit antibody, DNA-PK antibody, DNPK-1 antibody, DNAPK catalytic subunit antibody, DNPK 1 antibody, DNPK1 antibody, HYRC1 antibody, HYRC, P460, Hyper-Radiosensitivity Of Murine Scid Mutation Complementing 1, IMD26, XRCC7, P350

Rabbit Anti-DNA PKcs pT2609 affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids surrounding Thr 2609 of human DNA PKcs.

DNA PKcs pT2609 antibody is directed against the phosphorylated form of human DNA PKcs at the pT2609 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross-adsorbed against the non-phosphorylated form of the immunizing peptide. This phospho-specific polyclonal antibody reacts with phosphorylated pT2609 of human DNA PKcs. Reactivity with non-phosphorylated human DNA PKcs is minimal. A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. However, cross-reactivity is expected with mouse, rat, dog, chicken and horse DNA PKcs based on a high degree of homology to the immunogen sequence. Cross-reactivity with DNA PKcs homologues from other sources has not been determined.

Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.

Anti-DNA PKcs pT2609 antibody has been tested for use in ELISA, western blot, and IP. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 460 kDa in size corresponding to DNA PKcs by western blotting in the appropriate cell lysate or extract. Alternate splice variants have been described for this protein. Best western blotting results are seen when IP is performed prior to detection. This antibody detects an inducible signal at the correct height after DNA damage. However, the antibody may also detect an IR-inducible signal in cells lacking DNA PKcs (MO59J cells) at the same size as DNA PKcs if western blotting is performed directly. We believe that this additional band is 53BP1 that runs at the same size as DNA PKcs and is also phosphorylated in an IR-dependent manner. Similar results were seen with another phospho DNA PKcs antibody indicating that this result is general to antibodies to this phospho site, rather than specific to this antibody.