|DNA-PKcs (E6U3A) Rabbit mAb 38168||20 µl||
|Phospho-DNA-PKcs (Ser2056) (E9J4G) Rabbit mAb 68716||20 µl||
|Ku70 (D10A7) Rabbit mAb 4588||20 µl||
||H M R Mk||70||Rabbit IgG|
|Ku80 (C48E7) Rabbit mAb 2180||20 µl||
||H Mk||86||Rabbit IgG|
|DNA Ligase IV (D5N5N) Rabbit mAb 14649||20 µl||
|XLF Antibody 2854||20 µl||
|Artemis (D7O8V) Rabbit mAb 13381||20 µl||
||H Mk||90||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Pro608 of human DNA-PKcs protein, Val294 of mouse Ku70 protein, the carboxy terminus of human Ku80 protein, Leu771 of human DNA ligase IV protein, and Pro367 of human artemis protein. Phosphorylation-specific monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2056 of human DNA-PKcs protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the carboxy terminus of human XLF protein. Polyclonal antibodies are purified by peptide affinity chromatography.
DNA double-strand breaks (DSBs) are potentially hazardous lesions that can be induced by ionizing radiation (IR), radiomimetic chemicals, or DNA replication inhibitors. Cells recognize and repair DSBs via two distinct but partly overlapping signaling pathways, non-homologous end joining (NHEJ) and homologous recombination (HR). DNA repair via the HR pathway is restricted to S and G2 phases of the cell cycle, while NHEJ can occur during any phase. NHEJ machinery is also utilized in V(D)J recombination, a process that generates diversity in immunoglobulin and T cell receptor genes. Defects in both pathways have been associated with human disease, including cancer (1).
DNA repair through the NHEJ pathway involves a core group of proteins that includes the Ku heterodimer (Ku70/Ku80), DNA-PKcs, DNA ligase IV, XRCC4, and XLF. XLF interacts with XRCC4 and promotes the ligation of DNA strands by DNA ligase IV and the ligase cofactor XRCC4. The ATP-dependent ligation of free DNA ends is the final step in the NHEJ repair pathway (2). DNA ligase IV and the endonuclease artemis suppress homologous recombination and promote NHEJ (3).
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