Homologous Recombination (HR) DNA Repair Antibody Sampler Kit #99891
Product Information
Kit Usage Information
Protocols
- 2873: Western Blotting
- 3001: Western Blotting
- 7074: Western Blotting
- 8875: Western Blotting, Immunoprecipitation (Agarose)
- 9201: Western Blotting
- 10741: Western Blotting
- 13050: Western Blotting
- 14823: Western Blotting, Immunoprecipitation (Magnetic)
- 14956: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence
- 15016: Western Blotting, Immunoprecipitation (Magnetic)
Product Description
Specificity / Sensitivity
Each antibody in the Homologous Recombination (HR) DNA Repair Antibody Sampler Kit detects endogenous levels of its target protein. Phospho-ATM (Ser1981) (D25E5) Rabbit mAb recognizes endogenous levels of ATM protein only when phosphorylated at Ser1981. Phospho-p95/NBS1 (Ser343) Antibody detects endogenous levels of p95/NBS1 protein only when phosphorylated at Ser343.
Source / Purification
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.
Background
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).
- Hartlerode, A.J. and Scully, R. (2009) Biochem J 423, 157-68.
- Lee, J.H. and Paull, T.T. (2007) Oncogene 26, 7741-8.
- Sung, P. et al. (2003) J Biol Chem 278, 42729-32.
- Tutt, A. and Ashworth, A. (2002) Trends Mol Med 8, 571-6.
- Wang, H. et al. (2013) PLoS Genet 9, e1003277.
- Wen, J. et al. (2013) Oncogene 32, 4448-56.
Limited Uses
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