ELISA, Western Blotting (WB), Immunoprecipitation (IP)
Cross-Reactivity
Chimpanzee
Characteristics
Concentration Definition: by UV absorbance at 280 nm
Immunogen
This 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.
This affinity purified antibody has been tested for use in ELISA and by western blot. 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.
Restrictions
For Research Use only
Format
Liquid
Concentration
0.93 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C
Yoshikawa, Kashino, Ono, Watanabe: "Phosphorylated H2AX foci in tumor cells have no correlation with their radiation sensitivities." in: Journal of radiation research, Vol. 50, Issue 2, pp. 151-60, (2009) (PubMed).
Wanner, Mayer, Kehlbach, Rodemann, Dittmann: "Activation of protein kinase Cepsilon stimulates DNA-repair via epidermal growth factor receptor nuclear accumulation." in: Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology, Vol. 86, Issue 3, pp. 383-90, (2008) (PubMed).
Dittmann, Mayer, Kehlbach, Rodemann: "The radioprotector Bowman-Birk proteinase inhibitor stimulates DNA repair via epidermal growth factor receptor phosphorylation and nuclear transport." in: Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology, Vol. 86, Issue 3, pp. 375-82, (2008) (PubMed).
Dittmann, Mayer, Wanner, Kehlbach, Rodemann: "The radioprotector O-phospho-tyrosine stimulates DNA-repair via epidermal growth factor receptor- and DNA-dependent kinase phosphorylation." in: Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology, Vol. 84, Issue 3, pp. 328-34, (2007) (PubMed).
Dittmann, Mayer, Ohneseit, Raju, Andratschke, Milas, Rodemann: "Celecoxib induced tumor cell radiosensitization by inhibiting radiation induced nuclear EGFR transport and DNA-repair: a COX-2 independent mechanism." in: International journal of radiation oncology, biology, physics, Vol. 70, Issue 1, pp. 203-12, (2007) (PubMed).
DNA dependent Protein Kinase (also called DNAPK, DNPK1, HYRC1, Protein Kinase DNA Activated Catalytic Polypeptide, XRCC7 and p460) consists of the 460 kDa DNA PKcs and a heterodimeric regulatory complex comprised of p70 Ku and p80 Ku (Ku autoantigen). DNA PKcs is a nuclear protein serine/threonine kinase present in a wide variety of eukaryotic species. DNA PKcs phosphorylates transcription factors, Sp1, Oct-1, p53 and SV40 large T antigen. DNA PKcs is involved in repairing double stranded DNA breaks. At the onset of apoptosis, DNA PKcs is rapidly inactivated by cleavage of the catalytic subunit into smaller polypeptides. Proteolysis of DNA PKcs is inhibited by the cysteine protease inhibitors iodoacetamide and N-ethylmaleimide. Alternative splicing can occur for this protein to produce at least two isoforms. Synonyms: DNA dependent protein kinase catalytic subunit antibody, DNA PK catalytic subunit antibody, DNAPK antibody, DNAPK catalytic subunit antibody, DNPK 1 antibody, DNPK1 antibody, HYRC 1 antibody