Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101

akt   Akt S473   akt1   akt2   akt3   control slides   IHC   p-Akt   pAkt   phospho Akt   phospho-Akt   PKB   S473   SignalSlideTM  

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8101S 1 Pack ( 5 slides ) ¥1,680.00 现货查询 购买询价
8101 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Description

Each control slide contains formalin fixed, paraffin-embedded LNCaP cells, both untreated and treated with PI3-Kinase inhibitor LY294002, that serve as a control for Phospho-Akt (Ser473) immunostaining. Western blot analysis was performed on extracts derived from the same cells to verify the efficacy of the LY942002 treatment.

每一组对照包含福尔马林固定和石蜡包埋的, PI3K抑制剂LY294002处理及未处理的LNCaP细胞,它们作为Phospho-Akt (Ser473)免疫染色的对照组。Western被用来分析从相同细胞得到的提取物以验证LY942002 处理的效率。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded LNCaP cells untreated (left) or treated with LY294002 (right) using Phospho-Akt (Ser473) (736E11) Rabbit mAb (IHC Specific) #3787.

免疫组化分析石蜡包埋的LNCaP细胞,未处理组(左)和 LY294002 (右)处理组,所用抗体为Phospho-Akt (Ser473) (736E11) Rabbit mAb 兔单抗(IHC Specific) #3787。

Western Blotting

Western Blotting

Western blot analysis of extracts from LNCaP cells, untreated or treated with LY294002, using Akt Antibody #9272 (upper) orPhospho-Akt (Ser 473) (193H121) Rabbit mAb #4058 (lower). This assay serves as a control for the efficacy of the LY94002 treatment.

western blot 分析LNCaP细胞提取物,未处理组和LY294002处理组,所用抗体为Akt Antibody #9272 (上) 和Phospho-Akt (Ser 473) (193H121) Rabbit mAb兔单抗 #4058 (下). 本方法作为 LY94002 处理效率的对照。

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded LNCaP cells, untreated (left) or treated with LY294002 (right) using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060.

免疫组化分析石蜡包埋的LNCaP细胞,未处理组(左)和 LY294002处理组(右),所用抗体为Phospho-Akt (Ser473) (D9E) Rabbit mAb 兔单抗#4060。

Schematic

Schematic

Applications

These slides are intended for use in immunohistochemical assays. Please see the Companion Products for the products that can be used with these slides. 这些玻片专门应用于免疫组化检测中。能与它们共同使用的产品请参见配套产品。

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 (4)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。 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.

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Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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