Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-PLK1 (Thr210) (D5H7) Rabbit mAb #9062

No. Size Price
9062S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9062 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 62 Rabbit IgG
IP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

Phospho-PLK1 (Thr210) (D5H7) Rabbit mAb recognizes endogenous levels of PLK1 protein only when phosphorylated at Thr210.Phospho-PLK1 (Thr210) (D5H7) Rabbit mAb能够识别内源性苏氨酸(210位)磷酸化的PLK1蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr210 of human PLK1 protein.该单克隆抗体是由合成的人源的针对PLK1蛋白苏氨酸(210位)的肽段免疫动物生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, asynchronous or synchronized in mitosis, using Phospho-PLK1 (Thr210) (D5H7) Rabbit mAb (upper) or total PLK1 (208G4) Rabbit mAb #4513 (lower). Mitotic synchronization was performed by thymidine block followed by release into nocodazole and mitotic shake-off.Western blot方法检测Hela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) (D5H7) Rabbit mAb(上图)或总PLK1 (208G4) Rabbit mAb #4513 (下图)。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。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, asynchronous or synchronized in mitosis, using Phospho-PLK1 (Thr210) (D5H7) Rabbit mAb. The antibody was pre-incubated with a PLK1 phospho-Thr210 peptide or nonphospho-peptide as indicated. Mitotic synchronization was performed by thymidine block followed by release into nocodazole and mitotic shake-off.Western blot方法检测HeLa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) (D5H7) Rabbit mAb。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。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。

Background

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). 
 Substitution of Thr210 with Asp has been reported to elevate PLK1 kinase activity and delay/arrest cells in mitosis, while a Ser137Asp substitution leads to S-phase arrest (12). In addition, while DNA damage has been found to inhibit PLK1 kinase activity, the Thr210Asp mutant is resistant to this inhibition (13). PLK1 has been reported to be phosphorylated in vivo at Ser137 and Thr210 in mitosis; DNA damage prevents phosphorylation at these sites (14).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)。据报道,苏氨酸(210位)被天冬氨酸取代后可以提高PLK1的激酶活性并且使细胞延迟或停滞于有丝分裂中,而丝氨酸(137位)为天冬氨酸取代后则导致细胞停滞于S期(12)。另外,尽管DNA损伤被发现能够抑制PLK1激酶活性,苏氨酸(210位)被天冬氨酸取代后能够抵制这种抑制作用 (13)。据报道,在体内有丝分裂过程中 PLK1丝氨酸(137位)和苏氨酸(210位)被磷酸化;而DNA损伤可以抑制这些位置的磷酸化(14)。

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  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.

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