Phospho-Akt Isoform Antibody Sampler Kit #10396

Kit Usage Information

Protocols

• 2938: Western Blotting, Immunoprecipitation (Magnetic)
• 3063: Western Blotting, Immunoprecipitation (Magnetic)
• 4060: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 4691: Western Blotting, Immunoprecipitation (Agarose), Immunohistochemistry (Paraffin), Immunofluorescence, Flow
• 7074: Western Blotting
• 8599: Western Blotting, Immunoprecipitation (Magnetic)
• 9018: Western Blotting, Immunoprecipitation (Magnetic), Immunofluorescence, Flow

Product Description

The Phospho-Akt Isoform Antibody Sampler Kit provides an economical means of detecting the activation of Akt family members using phospho-specific and control antibodies. The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.

Specificity / Sensitivity

Each antibody in this kit recognizes endogenous levels of its specific target protein. Akt (pan) (C67E7) Rabbit mAb does not cross-react with other related proteins. Akt1 (C73H10) Rabbit mAb does not cross-react with Akt2 or Akt3. Akt2 (D6G4) Rabbit mAb does not cross-react with Akt1 or Akt3. Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473. Phospho-Akt1 (Ser473) (D7F10) XP^® Rabbit mAb (Akt1 Specific) recognizes endogenous levels of Akt1 protein only when phosphorylated at Ser473. It does not detect Akt2 protein when phosphorylated at Ser474. Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) recognizes endogenous levels of Akt2 protein only when phosphorylated at Ser474. This antibody does not cross-react with Akt1 protein when phosphorylated at Ser473 or with Akt3 protein when phosphorylated at Ser472.

Source / Purification

Phospho-Akt (Ser473) (D9E) XP^® Rabbit mAb is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt. Akt (pan) (C67E7) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of mouse Akt. Phospho-Akt1 (Ser473) (D7F10) XP^® Rabbit mAb (Akt1 Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser473 of human Akt1 protein. Akt1 (C73H10) Rabbit mAb is produced by immunizing animals with a synthetic peptide surrounding Leu110 of human Akt1. Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser474 of human Akt2 protein. Akt2 (D6G4) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues of human Akt2.

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).
There are three Akt isoforms (Akt1, Akt2 and Akt3) in mammals (20). Akt activation requires phosphorylation by mTORC2 at Ser473 of Akt1, Ser474 of Akt2, and Ser472 of Akt3 (20).

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. Manning, B.D. and Toker, A. (2017) Cell 169, 381-405.

Pathways & Proteins

Explore pathways + proteins related to this product.

Select Your Pathway/Protein AMPK Signaling Adherens Junction Dynamics Alzheimer's Disease Angiogenesis Apoptosis Regulation B Cell Receptor Signaling ESC Pluripotency and Differentiation ErbB/HER Signaling G1/S Checkpoint IL6 Signaling Inhibition of Apoptosis Insulin Receptor Signaling Microtubule Dynamics Mitochondrial Control of Apoptosis NF-kB Signaling PI3K/Akt Signaling Protein Kinase C Signaling T Cell Receptor Signaling TGF-ß Signaling Translation: eIF4E and p70S6K Warburg Effect mTOR Signaling Protein: P31749 Protein: P31751 Protein: Q9Y243

Database Links

UniProt ID: P31751 , Q9Y243 , P31749

Entrez-Gene Id: 208 , 10000 , 207

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