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Each lot of this DNA-PKcs antibody is quality control tested by Western blotting. Western blotting, suggested working dilution(s): Use 5 μg antibody per 5 ml antibody dilution buffer for each mini-gel. For immunofluorescence microscopy: Use a dilution range of 1-4 μg/ml. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
MOLT4 nuclear extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with anti-DNA-PKcs (Thr2609) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Lane 1, untreated MOLT4 cells. Lane 2, MOLT4 cells exposed to 2 Gy radiation. Lane 3, MOLT4 cells exposed to 10 Gy radiation. In both cases, cells were harvested 30 minutes after radiation.
DNA-protein kinase catalytic subunit (DNA-PKcs) also known as DNA-activated kinase is a nuclear 460-470 kD serine threonine kinase involved in double-stranded DNA break repair, VDJ recombination, and transcriptional modulation. DNA-PKcs must bind DNA ends to become active. DNA-PKcs is modified by phosphorylation and has been shown to interact with Ku70/Ku80, KIP, DNA-ligase IV, and XRCC4 proteins. Phosphorylated DNA-PKcs is upregulated after DNA damaage. The 10B1 monoclonal antibody recognizes phosphorylated human DNA-PKcs (Thr2609) and has been shown to be useful for immunofluorescence staining and Western blotting.
Other Names:
DNA-protein kinase catalytic subunit, DNA-activated protein kinase, DNA-dependent protein kinase
1. Hartley K, et al. 1995. Cell. 82:849. 2. Connelly M, et al. 1996. Gene. 175:271. 3. Douglas P, et al. 2002. Biochem. J. 368:243 4. Calsou P, et al. 2003. J. Mol. Biol. 326:93.
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Purified anti-DNA-PKcs Phosphorylated (Thr2609)
MOLT4 nuclear extract was resolved by electrophoresis, transferred to nitrocellulose, and probed with anti-DNA-PKcs (Thr2609) antibody. Proteins were visualized using a goat anti-mouse secondary conjugated to HRP and a chemiluminescence detection system. Lane 1, untreated MOLT4 cells. Lane 2, MOLT4 cells exposed to 2 Gy radiation. Lane 3, MOLT4 cells exposed to 10 Gy radiation. In both cases, cells were harvested 30 minutes after radiation.
