|ATM (D2E2) Rabbit mAb 2873||20 µl||
||H M||350||Rabbit IgG|
|Phospho-ATM (Ser1981) (D25E5) Rabbit mAb 13050||20 µl||
|Rad51 (D4B10) Rabbit mAb 8875||20 µl||
||H M R Mk||37||Rabbit IgG|
|BRCA1 (A8X9F) Rabbit mAb 14823||20 µl||
|BRCA2 (D9S6V) Rabbit mAb 10741||20 µl||
|Rad54 (D4W3Z) Rabbit mAb 15016||20 µl||
|p95/NBS1 (D6J5I) Rabbit mAb 14956||20 µl||
||H M R||95||Rabbit IgG|
|Phospho-p95/NBS1 (Ser343) Antibody 3001||20 µl||
|CtIP (D76F7) Rabbit mAb 9201||20 µl||
||H Mk||110||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
Monoclonal antibodies are produced by immunizing animals with recombinant human ATM, Rad51, BRCA1 and BRCA2, or synthetic peptides corresponding to residues surrounding Gly246 of human Rad54, Ala740 of human p95/NBS1, or residues near the carboxy terminus of human CtIP protein. Phosphorylation-specific monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser1981 of human ATM. Phosphorylation-specific polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to Ser343 of human p95/NBS1 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). DSBs that arise during S or G2 phase are repaired via HR, using the replicated sister chromatid as a repair template (1). Activation of ATM by autophosphorylation on Ser1981 occurs in response to exposed DNA DSBs. ATM regulates various responses to DNA damage, including regulation of HR factors (2). Rad51 recombinase polymerizes and forms a filament along single-stranded DNA, mediating HR with the help of auxiliary proteins, including Rad54 and BRCA2 (3). BRCA2 has been shown to be required for localization of Rad51 to sites of DSBs, and cells lacking BRCA1 and BRCA2 cannot repair DSBs through HR (4). NBS1 is critical for HR, and requires CDK-dependent association with CtIP and subsequent phosphorylation by ATM at Ser278 and Ser343 (5-6).
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