PathScan® Phospho-Chk1 (Ser317) Sandwich ELISA Kit #7870
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Supporting Data
| REACTIVITY | H |
Species Cross-Reactivity Key:
- H-Human
Product Information
Product Description
*Antibodies in this kit are custom formulations specific to kit.
Protocol
Specificity / Sensitivity
CST's PathScan® Phospho-Chk1 (Ser317) Sandwich ELISA Kit #7870 detects endogenous levels of Chk1 protein when phosphorylated at Ser317. As shown in Figure 1, a significant induction of Chk1 phosphorylation at Ser317 can be detected in HeLa cells following treatment with UV using the Phospho-Chk1 (Ser317) Sandwich ELISA Kit #7870. The levels of total Chk1 (phospho and nonphospho) remain unchanged as shown by Western analysis and by PathScan® Total Chk1 Sandwich ELISA Kit #7872 (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Human
Background
Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 by ATM/ATR, followed by autophosphorylation of Ser296. Activation occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).
- Liu, Q. et al. (2000) Genes Dev 14, 1448-59.
- Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
- Jiang, K. et al. (2003) J Biol Chem 278, 25207-17.
- Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16.
- Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97.
- Zeng, Y. et al. (1998) Nature 395, 507-10.
- Löffler, H. et al. (2006) Cell Cycle 5, 2543-7.
- Zachos, G. et al. (2007) Dev Cell 12, 247-60.
- Garber, K. (2005) J Natl Cancer Inst 97, 1026-8.
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