| 货号 | 5472T |
| 反应种属 | Human, |
| 来源宿主 | Rabbit |
| 应用 | WB |
| 目标/特异性 | Phospho-PLK1 (Thr210) Antibody detects endogenous levels PLK1 only when phosphorylated at threonine 210. |
| 使用方法 | WB(1:1000) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | At least 4 distinct polo-like kinases exist in mammalian cells: PLK1, PLK2, PLK3 and PLK4/SAK (1). PLK1 apparently plays many roles during mitosis, particularly in regulating mitotic entry and exit. The mitosis promoting factor (MPF), cdc2/cyclin B1, is activated by dephosphorylation of cdc2 (Thr14/Tyr15) by cdc25C. PLK1 phosphorylates cdc25C at Ser198 and cyclin B1 at Ser133 causing translocation of these proteins from the cytoplasm to the nucleus (2-5). PLK1 phosphorylation of Myt1 at Ser426 and Thr495 has been proposed to inactivate Myt1, one of the kinases known to phosphorylate cdc2 at Thr14/Tyr15 (6). Polo-like kinases also phosphorylate the cohesin subunit SCC1, causing cohesin displacement from chromosome arms that allow for proper cohesin localization to centromeres (7). Mitotic exit requires activation of the anaphase promoting complex (APC) (8), a ubiquitin ligase responsible for removal of cohesin at centromeres, and degradation of securin, cyclin A, cyclin B1, Aurora A and cdc20 (9). PLK1 phosphorylation of the APC subunits Apc1, cdc16 and cdc27 has been demonstrated in vitro and has been proposed as a mechanism by which mitotic exit is regulated (10,11).哺乳动物细胞中至少存在四种polo样激酶：PLK1，PLK2，PLK3和PLK4/SAK(1)。PLK1在有丝分裂中发挥多种作用，尤其是调节有丝分裂的进入和离开。有丝分裂促进因子(MPF)，cdc2/cyclin B1，通过cdc25C介导的cdc2去磷酸化而被激活。PLK1能够磷酸化cdc25C的第198位丝氨酸和周期蛋白B1的第133位丝氨酸，从而导致这些蛋白从细胞质向细胞核转移(2-5)。PLK1磷酸化Myt1的第426位丝氨酸和495位苏氨酸被认为能够失活Myt1，它是一种可以磷酸化cdc2第14位和15位苏氨酸的蛋白激酶(6)。Polo样蛋白激酶也可以磷酸化粘连蛋白亚基SCC1，引起染色体臂的粘连蛋白移位，促进粘连蛋白的中心体适当定位(7)。有丝分裂的退出需要激活后期促进复合物(APC)，一种负责中心体粘连蛋白清除的泛素连接酶并能够降解分离酶抑制蛋白，周期蛋白A，周期蛋白B1，极光激酶A和cdc20(9)。体外研究已表明PLK1磷酸化APC亚基Apc1，cdc16和cdc27，且被认为是一种调节有丝分裂退出的机制(10,11)。 |
| 存放说明 | -20C |
| 计算分子量 | 62 |
| 参考文献 | 1 . Nigg, E.A. (1998) Curr Opin Cell Biol 10, 776-83. 2 . Toyoshima-Morimoto, F. et al. (2002) EMBO Rep 3, 341-8. 3 . Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-20. 4 . Peter, M. et al. (2002) EMBO Rep 3, 551-6. 5 . Jackman, M. et al. (2003) Nat Cell Biol 5, 143-8. 6 . Nakajima, H. et al. (2003) J Biol Chem 278, 25277-80. 7 . Sumara, I. et al. (2002) Mol Cell 9, 515-25. 8 . Hauf, S. et al. (2001) Science 293, 1320-3. 9 . Peters, J.M. (1999) Exp. Cell Res. 248, 339-349. 10 . Kraft, C. et al. (2003) EMBO J 22, 6598-609. 11 . Kotani, S. et al. (1998) Mol Cell 1, 371-80. 12 . Jang, Y.J. et al. (2002) J Biol Chem 277, 44115-20. 13 . Smits, V.A. et al. (2000) Nat Cell Biol 2, 672-6. 14 . Tsvetkov, L. and Stern, D.F. (2005) Cell Cycle 4, 166-71. |
Western blot analysis of extracts from HT-29 cells, asynchronous or synchronized in mitosis using Phospho-PLK1 (Thr210) Antibody. The antibody was pre-incubated with a PLK1 phospho-Thr210 peptide or nonphospho-peptide as indicated. The same lysates were examined using total PLK1 (208G4) Rabbit mAb #4513.Mitotic synchronization was performed by thymidine block followed by release into nocodazole and mitotic shake-off.Western blot方法检测HT-29细胞提取物，分为非同步组和同步于有丝分裂期组，使用的抗体为Phospho-PLK1 (Thr210) Antibody。该抗体与苏氨酸（210位）磷酸化的PLK1多肽或特定的非磷酸化多肽预孵育。同一裂解液也使用总 PLK1 (208G4) Rabbit mAb #4513抗体检测。有丝分裂同步化是由胸腺嘧啶阻断后释放至诺考达唑从而使有丝分裂停止。 |
