9010:

Immunoprecipitation, Western Blotting

BRCA1 Antibody detects endogenous levels of total BRCA1 protein. Five human isoforms are produced by alternative splicing and alternative initiation. The nuclear isoforms 1, 2, and 4 are detected, whereas the cytoplasmic isoforms 3 and 5 are not. The antibody does not recognize BRCA2.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids near the amino terminus of human BRCA1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of untreated and UV-treated (50 mJ/cm2, 30 min) HeLa cells, using BRCA1 antibody.

The breast cancer susceptibility proteins BRCA1 and BRCA2 are frequently mutated in cases of hereditary breast and ovarian cancers and have roles in multiple processes related to DNA damage, repair, cell cycle progression, transcription, ubiquitination and apoptosis (1-4). BRCA2 has been shown to be required for localization of Rad51 to sites of double stranded breaks (DSBs) in DNA, and cells lacking BRCA1 and BRCA2 cannot repair DSBs through the Rad51-dependent process of homologous recombination (HR) (5). Numerous DNA-damage induced phosphorylation sites on BRCA1 have been identified, including serines 988, 1189, 1387, 1423, 1457, 1524 and 1542, and kinases activated in a cell cycle-dependent manner, including Aurora A and CDK2, can also phosphorylate BRCA1 at Ser308 and Ser1497, respectively (6-10). Cell cycle-dependent phosphorylation of BRCA2 at Ser3291 by CDKs has been proposed as a mechanism to switch off HR as cells progress beyond S-phase by blocking the carboxy-terminal Rad51 binding site (11).

1. Rahman, N. and Stratton, M.R. (1998) Annu. Rev. Genet. 32, 95-121.
2. Gayther, S. A. et al. (1999) Am. J. Hum. Genet. 65, 1021-1029.
3. Kerr, P. and Ashworth, A. (2001) Curr. Biol. 11, R668-R676.
4. Scully, R. and Livingston, D.M. (2000) Nature 408, 429-432.
5. Tutt, A. and Ashworth, A. (2002) Trends Mol. Med. 8, 571-576.
6. Okada, S. and Ouchi, T. (2003) J. Biol. Chem. 278, 2015-2020.
7. Cortez, D. et al. (1999) Science 286, 1162-1166.
8. Xu, B. et al. (2002) Cancer Res. 62, 4588-4591.
9. Ouchi, M. et al. (2004) J. Biol. Chem. 279, 19643-19648.
10. Ruffner, H. et al. (1999) Mol. Cell. Biol. 19, 4843-4854.
11. Esashi, F. et al. (2005) Nature 434, 598-604.

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• 4945 Phospho-FANCD2 (Ser222) Antibody
• 4526 Phospho-ATM (Ser1981) (10H11.E12) Mouse mAb
• 9012 BRCA2 Antibody
• 3425 Rad52 Antibody

For Research Use Only. Not For Use In Diagnostic Procedures.