Phospho-DNA-PK (Ser2056) Antibody

• 产品编号：CST-4215S      品牌：Cell Signaling Technology       原厂货号：4215S
• 产品分类：抗体 > 一抗 > 磷酸化抗体
• 应用分类：

描述：

 Phospho-DNA-PK (Ser2056) Antibody 能够识别内源性丝氨酸（2056位）磷酸化的DNA-PK蛋白。该多克隆抗体是由合成的人源的针对DNA-PK蛋白丝氨酸（2056位）的肽段免疫动物，采用蛋白A和多肽亲和层析技术纯化生产的。 DNA依赖性蛋白激酶(DNA-PK)是DNA双链断裂修复的一个重要因子。细胞缺少DNA-PK或DNA-PK被抑制都不能进行合适的非同源末端连接(NHEJ) (1-7)。DNA-PK由两个DNA结合亚基(Ku70 和Ku86)和一个450kDa的催化亚基(DNA-PKcs)组成(8)。据认为Ku70和Ku86组成的异源二聚体先于DNA-PKcs及其激活结合DNA双链断裂末端 (1, 9)。激活的DNA-PKcs是一种丝氨酸/苏氨酸激酶，能够体外激活一系列蛋白如p53，转录因子，RNA 聚合酶和Ku70/Ku86 (10, 11)。DNA-PKcs多位点的自体磷酸化，包括2609位苏氨酸，可导致DNA-PK激酶活性和NHEJ能力失活(12, 13)。然而，据发现DNA-PK在自体磷酸化之前可优先磷酸化其底物，表明DNA-PK可能在DAN修复机制中的解聚过程发挥作用(14, 15)。第2609位苏氨酸的磷酸化参与DNA-PK介导的双链断裂修复，而且磷酸化的DNA-PK伴随H2A.X和53BP1集中于DNA损伤位点(16)。DNA-PK蛋白丝氨酸（2056位）磷酸化是由双链DNA断裂和ATM激活引发的（17）。

1. Gottlieb, T.M. and Jackson, S.P. (1993) Cell 72, 131-42.
2. Hartley, K. O. et al. (1995) Cell 82, 840-856.
3. Rosenzweig, K.E. et al. (1997) Clin Cancer Res 3, 1149-56.
4. Jackson, S.P. and Jeggo, P.A. (1995) Trends Biochem Sci 20, 412-5.
5. Roth, D.B. et al. (1995) Curr Biol 5, 496-9.
6. Baumann, P. and West, S.C. (1998) Proc Natl Acad Sci U S A 95, 14066-70.
7. Chen, S. et al. (2001) J Biol Chem 276, 24323-30.
8. Jeggo, P.A. (1997) Mutat Res 384, 1-14.
9. Suwa, A. et al. (1994) Proc Natl Acad Sci U S A 91, 6904-8.
10. Anderson, C.W. and Lees-Miller, S.P. (1992) Crit Rev Eukaryot Gene Expr 2, 283-314.
11. Kuhn, A. et al. (1995) Genes Dev 9, 193-203.
12. Chan, D.W. and Lees-Miller, S.P. (1996) J Biol Chem 271, 8936-41.
13. Douglas, P. et al. (2002) Biochem. J. 368, 243-251.
14. Lees-Miller, S.P. et al. (1992) Mol Cell Biol 12, 5041-9.
15. Jackson, S.P. et al. (1990) Cell 63, 155-65.
16. Chan, D.W. et al. (2002) Genes Dev 16, 2333-8.
17. Yajima, H. et al. (2009) J Mol Biol 385, 800-10.

原厂资料：

Homology

Species predicted to react based on 100% sequence homology: Monkey

Specificity / Sensitivity

Phospho-DNA-PK (Ser2056) Antibody recognizes endogenous levels of DNA-PK protein only when phosphorylated at Ser2056.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser2056 of human DNA-PK protein. Antibodies are purified by protein A and peptide affinity chromatography.

Background

DNA-dependent protein kinase (DNA-PK) is an important factor in the repair of double-stranded breaks in DNA. Cells lacking DNA-PK or in which DNA-PK is inhibited fail to show proper nonhomologous end-joining (NHEJ) (1-7). DNA-PK is composed of two DNA-binding subunits (Ku70 and Ku86) and one 450 kDa catalytic subunit (DNA-PKcs) (8). It is thought that a heterodimer of Ku70 and Ku86 binds to double-stranded DNA broken ends before DNA-PKcs binds and is activated (1,9). Activated DNA-PKcs is a serine/threonine kinase that has been shown to phosphorylate a number of proteins in vitro, including p53, transcription factors, RNA polymerase, and Ku70/Ku86 (10,11). DNA-PKcs autophosphorylation at multiple sites, including Thr2609 and Ser2056, results in an inactivation of DNA-PK kinase activity and NHEJ ability (12,13). It has been demonstrated, however, that DNA-PK preferentially phosphorylates substrates before it autophosphorylates, suggesting that DNA-PK autophosphorylation may play a role in disassembly of the DNA repair machinery (14,15). Autophosphorylation at Thr2609 has also been shown to be required for DNA-PK-mediated double strand break repair, and phosphorylated DNA-PK co-localizes with H2A.X and 53BP1 at sites of DNA damage (16). Phosphorylation at Ser2056 occurs in response to double-stranded DNA breaks and ATM activation (17)

1. Gottlieb, T.M. and Jackson, S.P. (1993) Cell 72, 131-42.
2. Hartley, K. O. et al. (1995) Cell 82, 840-856.
3. Rosenzweig, K.E. et al. (1997) Clin Cancer Res 3, 1149-56.
4. Jackson, S.P. and Jeggo, P.A. (1995) Trends Biochem Sci 20, 412-5.
5. Roth, D.B. et al. (1995) Curr Biol 5, 496-9.
6. Baumann, P. and West, S.C. (1998) Proc Natl Acad Sci U S A 95, 14066-70.
7. Chen, S. et al. (2001) J Biol Chem 276, 24323-30.
8. Jeggo, P.A. (1997) Mutat Res 384, 1-14.
9. Suwa, A. et al. (1994) Proc Natl Acad Sci U S A 91, 6904-8.
10. Anderson, C.W. and Lees-Miller, S.P. (1992) Crit Rev Eukaryot Gene Expr 2, 283-314.
11. Kuhn, A. et al. (1995) Genes Dev 9, 193-203.
12. Chan, D.W. and Lees-Miller, S.P. (1996) J Biol Chem 271, 8936-41.
13. Douglas, P. et al. (2002) Biochem. J. 368, 243-251.
14. Lees-Miller, S.P. et al. (1992) Mol Cell Biol 12, 5041-9.
15. Jackson, S.P. et al. (1990) Cell 63, 155-65.
16. Chan, D.W. et al. (2002) Genes Dev 16, 2333-8.
17. Yajima, H. et al. (2009) J Mol Biol 385, 800-10.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibody.