Product Pathways - PI3K / Akt Signaling

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Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb #4060

Applications	Reactivity	Sensitivity	MW (kDa)	Isotype
W IP IHC-P IHC-F IF-IC F	H M R Hm Mk Dm Z B (C) (X) (Dg) (Pg)	Endogenous	60	Rabbit IgG

Applications Key: W=Western Blotting IP=Immunoprecipitation IHC-P=Immunohistochemistry (Paraffin) IHC-F=Immunohistochemistry (Frozen) IF-IC=Immunofluorescence (Immunocytochemistry) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey C=Chicken Dm=D. melanogaster X=Xenopus Z=Zebrafish B=Bovine Dg=Dog Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

4060:

Flow , IHC / Frozen* , IHC / Paraffin , Immunofluorescence , Immunoprecipitation , Western Blotting

* Product-specific protocol.

Specificity / Sensitivity

Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt.

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).

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.

Application References

• Gray, M.J. et al. (2008) J Natl Cancer Inst 100, 109-20. Applications: Western Blotting
• Fos, C. et al. (2008) J Immunol 181, 1969-77. Applications: Western Blotting
• Engelman, J.A. et al. (2008) Nat Med 14, 1351-6. Applications: IHC-P (paraffin)
• Guertin, D.A. et al. (2009) Cancer Cell 15, 148-159. Applications: IHC-P (paraffin) Western Blotting
• Allard, D. et al. (2008) J Biol Chem 283, 19739-47. Applications: IHC-P (paraffin)
• De Raedt, T. et al. (2011) Cancer Cell 20, 400-13. Applications: Western Blotting
• Kleiman, L.B. et al. (2011) Mol Cell 43, 723-37. Applications: Western Blotting
• Sykes, S.M. et al. (2011) Cell 146, 697-708. Applications: Flow Cytometry
• Cheung, R. et al. (2011) J Clin Invest 121, 4446-61. Applications: Western Blotting
• Siemens, N. et al. (2011) J Biol Chem 286, 21612-22. Applications: Western Blotting
• Lan, R. et al. (2012) Am J Physiol Renal Physiol , . Applications: IHC-P (paraffin)
• Matsuura, S. et al. (2012) Blood , . Applications: Western Blotting

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know !

Companion Products

• 4691 Akt (pan) (C67E7) Rabbit mAb
• 2965 Phospho-Akt (Thr308) (C31E5E) Rabbit mAb
• 8112 SignalStain^® Antibody Diluent
• 1140 Phospho-Akt (Ser473) Blocking Peptide
• 2967 Akt1 (2H10) Mouse mAb
• 4051 Phospho-Akt (Ser473) (587F11) Mouse mAb
• 2964 Akt2 (5B5) Rabbit mAb
• 3788 Akt3 (62A8) Rabbit mAb
• 9272 Akt Antibody
• 9916 Phospho-Akt Pathway Antibody Sampler Kit
• 8101 SignalSlide^® Phospho-Akt (Ser473) IHC Controls
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 4071 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (Alexa Fluor^® 488 Conjugate)
• 4075 Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb (Alexa Fluor^® 647 Conjugate)
• 8114 SignalStain^® Boost IHC Detection Reagent (HRP, Rabbit)

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For Research Use Only. Not For Use In Diagnostic Procedures.一、什么是XMT™ & XP™？

XMT™ (eXceptional Monoclonal Technology™)是一种开发单克隆抗体的技术（eXceptional Monoclonal Technology™中文直译为卓越的单克隆抗体技术），CST科学家就是使用XMT™技术开发的XP™单克隆抗体。XP™ (eXceptional Performance™)是CST使用XMT™技术开发出来的抗体（eXceptional Performance™中文直译为卓越的性能）。

二、XP™抗体的特点
XP™抗体较常规的单克隆抗体技术生产的抗体，具有更高的特异性，灵敏度，批次间的稳定性和更加卓越的检测效果，目前所有的XP™抗体均为兔源的单抗。
相关产品技术细节及操作说明请登录CST公司网站http://www.cellsignal.com或http://www.cst-c.com.cn获取。

n更高的特异性

磷酸化检测容易出现假阳性，保证客户的实验结果是客观的。

n更高的灵敏度

磷酸化状态及内源性蛋白的表达率相对较低，使用XP™抗体可以检测低表达和珍贵的样品。另外，较普通的抗体，抗体的用量会降低，节省了成本。

n保证批次间的稳定性

如果碰到批次更换经常要进行条件的优化，XP™抗体只会新一批比老的批次质量更好，所以客户减少不必要的再摸索过程，省时间，省成本。

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同样的经费，得到更高的性价比！

CST科学家会验证每一批次的抗体，如果按照CST推荐的实验方法都会得到与CST网站上该产品同样的结果图！一样的花费，让客户更节约，更快速，得到更好的检测结果！