| 货号 | 12302T |
| 同种亚型 | Rabbit IgG |
| 反应种属 | Human/Mouse/Monkey |
| 来源宿主 | Rabbit |
| 应用 | W/IP/IF-IC |
| 目标/特异性 | Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb recognizes endogenous levels of Chk1 protein only when phosphorylated at Ser317. This antibody also detects an 80 kDa protein of unknown origin in some cell lines. |
| 使用方法 | WB(1:1000) IP (1:50) IF-IC (1:800) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | 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 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).Chk1激酶是ATM/ATR激酶的下游，在DNA损伤检验点控制，胚胎发育和癌症抑制过程中发挥了重要作用（1）。Chk1的激活包括317位和345位丝氨酸的磷酸化，这一过程是对DNA复制抑制和某些基因毒性压力的应激（2）。345位丝氨酸磷酸化可以帮助Chk1在检验点激活后定位到细胞核（3），317位磷酸化和PTEN某些特定位点磷酸化可以是细胞在停滞的DNA复制后重新进入细胞周期（4）。Chk1通过部分调控磷酸酯酶cdc25家族蛋白，发挥它对细胞周期检验点的调控作用。Chk1磷酸化cdc25A促进它的蛋白酶解并通过14-3-3结合抑制它的活性（5）。激活的Chk1可以通过磷酸化cdc25C使其失活，阻断cdc2的活性使其无法进入细胞周期（6）。中心体Chk1已经证明可以磷酸化cdc25B，也能抑制cdc25B对CDK1-cyclin B1的激活作用，从而废除有丝分裂纺锤体的形成和染色质缩合（7）。此外，Chk1通过调节aurora B和BubR1发挥调控纺锤体检验点的功能（8）。研究提示由于Chk1被抑制后会导致很多癌细胞死亡，Chk1可以作为癌症治疗的药物靶点（9）。 |
| 存放说明 | -20C |
| 计算分子量 | 56 |
| 参考文献 | 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. |
Confocal immunofluorescent analysis of HeLa cells, untreated (left), UV-treated (center), or UV and λ phosphatase-treated (right), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).对未处理(左)，UV处理(中)或UV和λ phosphatase处理(右)的HeLa细胞，使用Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb(绿色)进行激光共聚焦免疫荧光分析。肌动蛋白丝使用DY-554鬼笔环肽标记(红色)。 | |
Western blot analysis of extracts from 293 and NIH/3T3 cells, untreated (-) or UV-treated (100 mJ, 1 hr recovery; +), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb. The blot on the right was treated with calf intestinal phosphatase (CIP) before western blot.未处理(-)或UV处理(100 mJ, 1 hr 恢复; +)的293和NIH/3T3细胞提取物，使用Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb进行western blot分析。右边的位点在western blot前使用牛小肠碱性磷酸酶(CIP)处理。 | |
Immunoprecipitation of phospho-Chk1 (Ser317) from 293 cell extracts treated with UV (100 mJ, 1 hr recovery) using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (lane 2) or Rabbit (D1AG) mAb IgG XP® Isotype Control #3900 (lane 3). Lane 1 is 10% input.使用Phospho-Chk1 (Ser317) (D12H3) XP®Rabbit mAb (lane 2)或兔(D1AG) mAb IgG XP® Isotype Control #3900 (lane 3)对经过UV处理(100 mJ, 1 hr 恢复)的293细胞提取物中的磷酸化Chk1进行免疫沉淀实验。Lane 1是10%上样量。 |
