DNA PK, Catalytic Subunit, multiple clones (DNA-dependent Protein Kinase)

DNA-dependent Protein Kinase (DNA-PKcs) phophorylates certain transcription factors, including Sp1, Oct-1, p53 and SV40 large T antigen in vitro. In vivo. It is involved in repairing double strand DNA breaks.\n\nApplications: \nSuitable for use in Gel Supershift, Immunohistochemistry, Immunofluorescence, Immunoprecipitation and Western Blot. Other applications not tested.\n\nRecommended Dilution:\nImmunoprecipitation (Native and denatured): 2ug/mg protein lysate (Use Protein G).\nWestern Blot: 1-2ug/ml.\nImmunohistochemistry (Formalin/pariffin): 2-4ug/ml for 30 min at RT. Note: Stainig of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6 for 10-20minutes followed by cooling at RT for 20 minutes.\nGel Supershift: Use antibody at 1mg/ml\nOptimal dilutions to be determined by the researcher.\n\nPositive Control: \nLS174T cells. Tonsil or testis.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

D3949-20

Size

500ul

Applications

IF, IHC, IP, WB

Hosts

Mouse

Reactivities

Hum, Mouse, Ch/Bird, Xen/Amph

Form

Supplied as a liquid in PBS, pH 7.4, 0.2% BSA, 0.1% sodium azide before the addition of glycerol to 40%. Also available without BSA and azide. See D3949-20X.

P Type

Mab

Purity

Purified by Protein A/G affinity chromatography.

Isotype

IgG1+IgG2a

References

1. Carter T, et. al. Molecular and Cellular Biology, 1990, 10(12):6460-71.1. Anderson, CW and TH Carter. 1996. The DNA-Activated Protein Kinase, DNA-PK. in, Current topics in Microbiology and Immunology , R. Jessberger, ed., Springer-Verlag. 217:91-111. 2. Bannister AJ, TM Gottlieb, T Kouzarides, and SP Jackson. 1993. c-Jun is phosphorylated by the DNA-dependent protein kinase: in vitro definition of the minimal kinase recognition motif. Nucleic Acid Res. 21:1289-1295. 3. Biedermann, KA, J Sun, AJ Giaccia, LM Tosto, and JM Brown. 1991. Scid mutation in mice confers hypersensitivity to ionizing radiation and a deficiency in DNA double stranded break repair. Proc. Natl. Acad. Sci. USA. 88:1394-1397. 4. Blier, PR, AJ Griffith, J Craft, and JA Hardin. 1993. Binding of Ku protein to DNA. Measurement of affinity for ends and demonstration of binding to nicks. J Biol. Chem. 268:7594-7601. 5. Blunt, T, NJ Finnie, GE Taccioli, GCM Smith, J Demengeot, TM Gottlieb, R Mizuta, AJ Varghese, FW Alt, PA Jeggo, and SP Jackson. 1995. Defective DNA dependent protein kinase activity is linked to V(D)J recombination and DNA repair defects associated with the murine scid mutation. Cell 80:813-823. 6. Brush, GS, CW Anderson, and TJ Kelly. 1994. The DNA activated protein kinase is required for the phosphorylation of replication protein A during simian virus 40 DNA replication. Proc. Natl. Acad. Sci. USA. 91:12520-12524. 7. Cai Q-Q, A Plet, J Imbert, M Lafage-Pochitaloff, C Cerdan, and J-M Blanchard. 1994. Chromosomal location and expression of the genes coding for Ku p70 and p80 in human cell lines and normal tissues. Cytogenet. Cell. Genet. 65:221-227. 8. Carter, T, I Vancurov

Additional Info

Recognizes the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) at 460kD. Species Crossreactivity: human, mouse, hamster, Xenopus and chicken.