Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-Akt (Thr308) (C31E5E) Rabbit mAb #2965

Akt T308   akt1   akt2   akt3   p-Akt   pAkt   phospho Akt   phospho-Akt   PKB   sc-135650   T308  

No. Size Price
2965L 300 µl ( 30 western blots ) ¥9,059.00 现货查询 购买询价
2965S 100 µl ( 10 western blots ) ¥3,780.00 现货查询 购买询价
2965 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Hamster,Monkey, Endogenous 60 Rabbit IgG
F 1:100
IF-IC 1:1600

Species cross-reactivity is determined by western blot.

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

Specificity / Sensitivity

Phospho-Akt (Thr308) (C31E5E) Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Thr308.

Phospho-Akt (Thr308) (C31E5E) Rabbit mAb能够检测内源性的Thr308位点磷酸化的AKT蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Thr308 of mouse Akt.

该单克隆抗体是采用合成的与鼠源AKT蛋白Thr308位点周围序列相对应的磷酸肽免疫动物而生产的。

IF-IC

IF-IC

Confocal immunofluorescent analysis of C2C12 cells, insulin-treated (left) or LY294002-treated (right), using Phospho-Akt (Thr308) (C31E5E) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

激光共聚焦荧光法检测C2C12 细胞, 左图为胰岛素处理的 ,右图为LY294002处理的,检测抗体为Phospho-Akt (Thr308) (C31E5E) Rabbit mAb兔单抗,呈绿色。微丝用Alexa Fluor® 555 phalloidin标记(红色)。蓝色伪彩是 DRAQ5® #4084 (fluorescent DNA dye)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue), using Phospho-Akt (Thr308) (C31E5E) Rabbit mAb.

流式细胞技术分析Jurkat细胞,未处理(绿色), LY294002, Wortmannin 和 U0126处理(蓝色);使用的抗体是Phospho-Akt (Thr308) (C31E5E) Rabbit mAb兔单抗。

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 and Jurkat cells, untreated, PDGF-treated or LY294002-treated as indicated, using Phospho-Akt (Thr308) (C31E5E) Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

western blot方法检测细胞提取物:未处理和PDGF, LY294002处理的NIH/3T3 和 Jurkat细胞,使用的抗体为 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb兔单抗 (上图) 和 Akt (pan) (C67E7)Rabbit mAb 兔单抗#4691 (下图).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of serum-starved NIH/3T3 cells, untreated (blue) or treated with mouse platelet-derived growth factor BB (200ng/ml, 10 min; green), using Phospho-Akt (Thr308) (D31E5E) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412.

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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