Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-Akt (Ser473) Antibody #9271

Akt S473   AKT1   akt2   akt3   p-akt   pakt   phospho akt   phospho-Akt   PKB   RAC   RAC-PK   RAC-PK alpha   sc-7985  

No. Size Price
9271L 300 µl ( 30 western blots ) ¥8,792.00 现货查询 购买询价
9271S 100 µl ( 10 western blots ) ¥3,780.00 现货查询 购买询价
9271T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
9271 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Hamster,D. melanogaster,Bovine,Dog,Pig, Endogenous 60 Rabbit
IP 1:100
F 1:50
IF-IC 1:25

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Monkey, Chicken, Xenopus, Horse,

Specificity / Sensitivity

Phospho-Akt (Ser473) Antibody detects endogenous levels of Akt1 only when phosphorylated at Ser473. This antibody also recognizes Akt2 and Akt3 when phosphorylated at the corresponding residues. It does not recognize Akt phosphorylated at other sites, nor does it recognize phosphorylated forms of related kinases such as PKC or p70 S6 kinase.

Phospho-Akt (Ser473) Antibody可以检测Ser473 位点磷酸化的Akt1。其也识别在Akt2和Akt3相应位点磷酸化的蛋白。但并不识别Akt其它位点磷酸化的蛋白以及其它相关激酶如PKC或p70 S6的磷酸化形式。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser473 of mouse Akt. Antibodies are purified by protein A and peptide affinity chromatography.

多克隆抗体是采用合成的与小鼠AktSer473位点周围序列相对应的磷酸肽免疫动物生产的。抗体采用蛋白A和肽亲和层析法纯化。

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF, wortmannin, LY294002, rapamycin or PD98059, using Phospho-Akt (Ser473) Antibody.

Western blot 分析NIH/3T3 细胞提取物, 非处理组和PDGF, wortmannin, LY294002, rapamycin 或PD98059处理组,所用抗体为 Phospho-Akt (Ser473) Antibody。

Western Blotting

Western Blotting

Western blot analysis of immunoprecipitated Akt from 293 cells transiently transfected with HA-tagged Akt (WT), HA-tagged K179A mutant Akt and HA-tagged K179A/S473A mutant Akt, using Phospho-Akt (Ser473) Antibody (upper), Akt antibody (middle) or HA antibody (lower). Phospho-Akt (Ser473) Antibody does not recognize Akt with an alanine substituion at Ser473. (Polakiewicz, R.D. et al. [1998] J. Biol. Chem. 273, 23534-23541.)

Western blot 分析瞬时转染HA-tagged Akt (WT), HA-tagged K179A突变Akt 和 HA-tagged K179A/S473A 突变 Akt的293 细胞免疫沉淀的Akt,所用抗体为Phospho-Akt (Ser473) Antibody (上), Akt antibody (中) or HA antibody (下)。 Phospho-Akt (Ser473) Antibody 不能识别丙氨酸代换的Akt。(Polakiewicz, R.D. et al. [1998] J. Biol. Chem. 273, 23534-23541.)

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF for the indicated times, using Phospho-Akt (Ser473) Antibody (upper) or Akt Antibody #9272 (lower).

Western blot分析NIH/3T3细胞的提取物,分为未处理组和PDGF处理组,如图中的时间所示,使用的抗体是Phospho-Akt (Ser473) Antibody (上) 或者 Akt Antibody #9272 (下)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of LNCaP cells, untreated (green) or LY294002-treated (blue), using Phospho-Akt (Ser473) Antibody compared to a nonspecific negative control antibody (red).

流式细胞术分析未处理(绿色)或者LY294002处理(蓝色)的LNCaP细胞,使用的抗体是Phospho-Akt (Ser473) Antibody,并和非特异性阴性对照抗体比较(红色)。

IF-IC

IF-IC

Confocal immunofluorescent images of C2C12 cells serum starved and treated with or without insulin as indicated and labeled with Phospho-Akt (Ser473) Antibody (red). Actin filaments have been labeled with fluorescein phalloidin.

免疫荧光共聚焦分析C2C12细胞,如图分别为血清饥饿,胰岛素未处理或者处理组,并应用Phospho-Akt (Ser473) 抗体标记(红色)。肌动蛋白纤维由荧光素-鬼笔环肽(fluorescein phalloidin)所标记。

Background

Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1/CIP1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).

Akt,又被称为PKB 或 Rac,在细胞的生长和凋亡中起到关键作用(1-3)。该蛋白激酶可以被胰岛素和多种生长和存活因子激活,在涉及PI3K激酶的wortmannin敏感信号通路中发挥作用(2,3)。Akt可由磷脂结合激活,该过程通过活化环中的Thr308位点以及羧基端Ser473位点的磷酸化完成,其中Thr308的磷酸化由PDK1完成(4)。 之前推测PDK2在Ser473位点磷酸化Akt,后被证实为哺乳动物rapamycin靶蛋白mTOR 的作用, 它存在在一个含有rictor和Sin1 的rapamycin非敏感复合体中。 Akt促进细胞的生长通过抑制细胞的凋亡 ,例如Akt可以抑制下游靶蛋白Bad (7), forkhead 转录因子 (8), c-Raf (9), and caspase-9。PTEN是PI3K/Akt信号通路的主要负调控因子(10)。 LY294002是特异性的 PI3K 激酶抑制剂(11)。Akt 的另外一个主要功能是通过磷酸化并进而抑制GSK-3α 和 β来调控糖原的合成(12,13)。 Akt 也可以调控胰岛素诱导的葡萄糖转运 (12)。除此之外,Akt还可以调控细胞周期,这个功能通过抑制GSK-3β,从而调控其下游的Cyclin D1 的磷酸化和降解 (14),或者负向调控cyclin依赖的激酶抑制因子 p27 Kip (15) 和 p21 Waf1/CIP1 (16)来实现。 Akt 也可以通过直接磷酸化含有raptor的rapamycin敏感复合体中的mTOR来调控细胞的生长(17)。 更重要的是, Akt磷酸化并失活TSC2, 而TCS2是mTOR-raptor复合物中mTOR的抑制因子(18,19)。

  1. Franke, T.F. et al. (1997) Cell 88, 435-7.
  2. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
  3. Franke, T.F. et al. (1995) Cell 81, 727-36.
  4. Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
  5. Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
  6. Jacinto, E. et al. (2006) Cell 127, 125-37.
  7. Cardone, M.H. et al. (1998) Science 282, 1318-21.
  8. Brunet, A. et al. (1999) Cell 96, 857-68.
  9. Zimmermann, S. and Moelling, K. (1999) Science 286, 1741-4.
  10. Cantley, L.C. and Neel, B.G. (1999) Proc Natl Acad Sci USA 96, 4240-5.
  11. Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
  12. Hajduch, E. et al. (2001) FEBS Lett 492, 199-203.
  13. Cross, D.A. et al. (1995) Nature 378, 785-9.
  14. Diehl, J.A. et al. (1998) Genes Dev 12, 3499-511.
  15. Gesbert, F. et al. (2000) J Biol Chem 275, 39223-30.
  16. Zhou, B.P. et al. (2001) Nat Cell Biol 3, 245-52.
  17. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
  18. Inoki, K. et al. (2002) Nat Cell Biol 4, 648-57.
  19. Manning, B.D. et al. (2002) Mol Cell 10, 151-62.
  20. Devi, L. and Ohno, M. (2015) Transl Psychiatry 5, e562.

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