PathScan? Total Chk1 Sandwich ELISA Antibody Pair

• 产品编号：CST-7873S      品牌：CST       原厂货号：7873S
• 产品分类：ELISA及其它免疫分析 > ELISA > ELISA检测抗体对
• 应用分类：

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描述：

关于该抗体对的特异性和敏感性，请参阅相应的PathScan® Sandwich ELISA试剂盒。注：经过内部测试，该抗体对能够检测指定物种的蛋白，但是也可以检测来自其他物种的同源蛋白。相对于CST的PathScan® Total Chk1 Sandwich ELISA Kit #7872，PathScan® Total Chk1 Sandwich ELISA Antibody Pair提供了一个经济的选择。我们提供了捕获和检测抗体(100X储备液)和HRP-结合的二级抗体(1000X储备液)，针对4 x 96 孔ELISAs的试剂也非常充足。Chk1 mouse capture antibody在含有PBS的96孔板中包被过夜。封闭后加入Chk1 detection antibody和 anti-rabbit IgG HRP conjugated antibody，随后加入细胞裂解液。加入HRP底物(TMB)进行显色。显色过程中450nm吸光度的强度和总Chk1的量成正比。该试剂盒中的抗体具有特异性的固定组成。 Chk1激酶能够充当ATM/ATR激酶的下游并在DNA损伤检验点控制、胚胎发育和肿瘤抑制中发挥重要作用(1)。Chk1的激活涉及317位和345位丝氨酸的磷酸化，其发生响应DNA复制阻滞和某些类型的基因毒性应激反应(2)。尽管345位丝氨酸的磷酸化有助于伴随检验点激活的Chk1定位到核，317位丝氨酸的磷酸化连同位点特异性的PTEN磷酸化允许DNA复制停滞引起的细胞周期折返(4)。Chk1发挥其细胞周期检验点调节机制部分经由调控磷酸酶家族的cdc25。Chk1磷酸化cdc25A后靶向蛋白降解并通过14-3-3的结合抑制自身的活性(5)。激活的chk1能够通过216位丝氨酸的磷酸化失活cdc25C，阻塞cdc2的激活和有丝分裂转换的进入(6)。中心体的chk1被认为可以磷酸化cdc25B并抑制其激活CDK1-cyclin B1，藉此废除有丝分裂纺锤体的形成和染色质凝聚(7)。此外，Chk1通过调节Aurora B和BubR1在纺锤体检验点功能中发挥重要作用(8)。鉴于Chk1的抑制作用能够引起许多肿瘤细胞系的死亡，它已经成为肿瘤治疗中心的药物靶点(9)。

1. Liu, Q. et al. (2000) Genes Dev 14, 1448-59.
2. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
3. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17.
4. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16.
5. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97.
6. Zeng, Y. et al. (1998) Nature 395, 507-10.
7. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7.
8. Zachos, G. et al. (2007) Dev Cell 12, 247-60.
9. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8.

原厂资料：

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species. 

Description

CST's PathScan® Total Chk1 Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total Chk1 Sandwich ELISA Kit #7872. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Chk1 mouse capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added, followed by Chk1 detection antibody and anti-rabbit IgG HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of total Chk1. 
 Antibodies in this kit are custom formulations specific to the kit.

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 and 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 reentry 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). Chk1 has emerged as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9). 

1. Liu, Q. et al. (2000) Genes Dev 14, 1448-59.
2. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
3. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17.
4. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16.
5. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97.
6. Zeng, Y. et al. (1998) Nature 395, 507-10.
7. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7.
8. Zachos, G. et al. (2007) Dev Cell 12, 247-60.
9. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8.

注意事项：

Storage: Chk1 capture antibody and Chk1 detection antibody are 
stored at 4ºC. Anti-Rabbit IgG-HRP Linked Antibody is stored at -20ºC.