Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Phospho-ENSA (Ser67)/ARPP19 (Ser62) Antibody #5240

No. Size Price
5240S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
5240 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Rat, Endogenous 15 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Homology

Species predicted to react based on 100% sequence homology: Mouse,

Specificity / Sensitivity

Phospho-ENSA (Ser67)/ARPP19 (Ser62) Antibody recognizes endogenous levels of ENSA and ARPP19 proteins only when phosphorylated at Ser67 and Ser62, respectively.Phospho-ENSA (Ser67)/ARPP19 (Ser62) Antibody能够分别检测内源性丝氨酸(67位)和丝氨酸(62位)磷酸化的ENSA蛋白和ARPP19蛋白。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser67/Ser62 of human ENSA/ARPP19 protein. Antibodies are purified by protein A and peptide affinity chromatography.该多克隆抗体是由合成的人员的针对ENSA/ARPP19蛋白丝氨酸(67位)和丝氨酸(62位)的磷酸化肽段免疫动物,采用蛋白A和多肽亲和层析技术纯化生产的。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or treated (+) with thymidine (2 mM, 16 hr) followed by release into nocodazole (100 ng/ml, 24 hr), using Phospho-ENSA (Ser67)/ ARPP19 (Ser62) Antibody (upper) or ENSA Antibody #8770 (lower).Western blot方法检测Hela细胞提取物,分为非处理组(-)和胸腺嘧啶处理(2 mM, 16小时)释放至诺考达唑(100 ng/ml, 24 小时)组,使用的抗体为Phospho-ENSA (Ser67)/ ARPP19 (Ser62) Antibody (上图) 或ENSA Antibody #8770 (下图).

Background

Mitotic control is important for normal growth, development, and maintenance of all eukaryotic cells. Research studies have demonstrated that inappropriate control of mitosis can lead to genomic instability and cancer (reviewed in 1,2). A regulator of mitosis, Greatwall kinase (Gwl), was first identified in Drosophila melanogaster (3). Subsequent studies showed that, based on sequence homology and function, microtubule-associated serine/threonine kinase-like (MASTL) is the human ortholog of Gwl (4). Regulation of MASTL/Gwl activation has been shown to be critical for the correct timing of mitosis. Research shows that Gwl is activated by hyperphosphorylation (5). The phosphorylation of human Gwl at Thr194 and Thr207 by active cyclin B1-cdc2 leads to possible autophosphorylation at Ser875 (Ser883 in Xenopus), which stabilizes the kinase. Activated Gwl then phosphorylates α-Endosulfine (ENSA) and cAMP-regulated phosphoprotein 19 (ARPP19) at Ser67 and Ser62, respectively. Phosphorylated ENSA and ARPP19 inhibit the activity of the B-55-subunit associated form of protein phosphatase 2A (PP2A-B55), allowing for complete phosphorylation of mitotic substrates by cyclin B-cdc2 and mitotic entry. When MASTL/Gwl is inactivated, PP2A-B55 reactivates, which leads to dephosphorylation of cyclin B-cdc2 and mitotic exit (5,6, reviewed in 7).有丝分裂控制对于所有真核细胞的正常生长、发育和维护是非常重要的。研究表明,有丝分裂控制不当可导致基因组不稳定和癌症(1,2中已论述)。Greatwal l激酶(GWL),有丝分裂的一个调节子,是首次发现于黑腹果蝇中(3)。随后的研究表明,基于序列同源性和功能,微管相关的丝氨酸/苏氨酸激酶样(MASTL)是人类GWL的直接同源物(4)。 MASTL/ GWL激活的调节已被证明对于有丝分裂的正确时序是至关重要的。研究表明,GWL通过度磷酸化被激活(5)。人GWL的Thr194和Thr207经由活性细胞周期素B1-cdc2磷酸化可导致适当的Ser875(在非洲爪蟾为Ser883)自身磷酸化,能够稳定激酶。活性的GWL然后分别在Ser67和Ser62磷酸化α-Endosulfine(ENSA)和环磷酸腺苷调节的磷酸化蛋白19(ARPP19)。磷酸化的ENSA和ARPP19能够抑制B-55-亚单位的活性以及联合形式的蛋白磷酸酶2A(PP2A-B55),允许细胞周期蛋白B-cdc2介导的有丝分裂底物完整磷酸化和进入有丝分裂。当MASTL/ GWL被灭活,PP2A-B55重新激活,从而导致细胞周期蛋白B-cdc2的去磷酸化和有丝分裂退出(5,6,7中已论述)。

  1. Eichhorn, P.J. et al. (2009) Biochim Biophys Acta 1795, 1-15.
  2. Norbury, C. and Nurse, P. (1992) Annu Rev Biochem 61, 441-70.
  3. Yu, J. et al. (2004) J Cell Biol 164, 487-92.
  4. Voets, E. and Wolthuis, R.M. (2010) Cell Cycle 9, 3591-601.
  5. Blake-Hodek, K.A. et al. (2012) Mol Cell Biol 32, 1337-53.
  6. Vigneron, S. et al. (2011) Mol Cell Biol 31, 2262-75.
  7. Lorca, T. and Castro, A. (2012) Oncogene 32, 537-543.

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