Akt?Alexa Fluor? 488 Conjugated Antibody Sampler Kit

• 产品编号：CST-12046S      品牌：Cell Signaling Technology       原厂货号：12046S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

该抗体能识别内源性水平的在Ser473磷酸化的Akt。Phospho-Akt (Thr308) (C31E5E) Rabbit mAb (Alexa Fluor® 488 Conjugate)能识别内源性水平的在Thr308磷酸化的Akt。Akt (pan) (C67E7) Rabbit mAb (Alexa Fluor® 488 Conjugate)能识别内源性水平的总Akt蛋白。单克隆抗体通过用合成的磷酸肽或合成肽免疫动物制备。该磷酸肽是人Akt蛋白Ser473周围的残基、小鼠Akt蛋白Thr308周围的残基。该合成肽是小鼠Akt蛋白羧基端附近的残基。该试剂盒提供了一种经济的手段来研究Akt的活化状态，不需要荧光二抗。Akt，也被称为PKB或Rac，在控制存活和凋亡中起着至关重要的作用（1-3）。这种蛋白激酶由胰岛素和许多生长和存活因子活化，在渥曼青霉素敏感的通路（包括PI3激酶）中起作用（2,3）。Akt可以被磷脂结合和激活环在Thr308位点被PDK1磷酸化激活（4），也可以被羧基端的Ser473位点磷酸化激活。先前难以捉摸的PDK2能引起Akt Ser473位点的磷酸化，它已经和rictor及Sin1一起被确定是钠巴霉素不敏感复合物中钠巴霉素的哺乳动物靶子（mTOR）（5,6）。Akt通过磷酸化作用和使数个靶点（包括Bad（7）、叉头转录因子（8）、c-Raf (9)和caspase-9）失活来抑制凋亡，从而提高细胞存活。PTEN磷酸酶是一种主要的PI3激酶/Akt信号通路的负调节剂（10）。LY294002是一种特异的PI3激酶抑制剂（11）。Akt另一个重要功能是通过磷酸化作用及使GSK-3α和β失活来调节糖原合成（12,13）。Akt可能对胰岛素刺激的葡萄糖转运也有作用（12）。除了在存活和糖原合成方面的作用，Akt还通过阻止GSK-3β介导的磷酸化作用和cyclin D1的降解及负调节CDK抑制蛋白p27 Kip1 (15)和p21 Waf1/Cip1 (16)来影响细胞周期调节（14）。Akt能直接使钠巴霉素敏感复合物（包括raptor）中的mTOR磷酸化，因而在细胞生长方面也有至关重要的作用(17)。更重要的是，Akt能磷酸化和灭活抗结核菌素(TSC2)，一种mTOR-raptor复合物中的mTOR抑制剂(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.

原厂资料：

Specificity / Sensitivity

Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of Akt only when phosphorylated at Ser473. Phospho-Akt (Thr308) (C31E5E) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of Akt only when phosphorylated at Thr308. Akt (pan) (C67E7) Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total Akt protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt protein,a synthetic phosphopeptide corresponding to residues around Thr308 of mouse Akt protein, or a synthetic peptide corresponding to residues near the carboxy terminus of mouse Akt protein.

Description

The Akt Alexa Fluor® 488 Conjugated Antibody Sampler Kit provides an economical means to study the activation state of Akt without the need for a fluorescent secondary antibody.

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 Kip1 (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.

注意事项：

Storage: Supplied in PBS (pH 7.2), less than 0.1% sodium
azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibod-
ies. Protect from light. Do not freeze.