Technical Data
Akt Pathway, phosphorylated BioAssay™ Sample Kit
Kits and Assays Storage: -20°CShipping: Blue Ice
Synthetic phosphopeptides corresponding to residues surrounding Ser473 or Thr308 of mouse Akt, Ser9 of human GSK-3beta, Ser256 of human FKHR, Ser380 of human PTEN or Ser241 of human PDK1. Each antibody recognizes the phosphorylated form of its specific target.

Akt, also referred to as PKB or Rac, plays a critical role in controlling the balance between survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors, and functions 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 also by phosphorylation within the C-terminus at Ser473. Akt functions to promote cell survival by inhibiting apoptosis by means of its ability to phosphorylate and inactivate several targets, including Bad (5), Forkhead transcription factors (6) and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (7). LY294002 is a specific PI3 kinase inhibitor (8).

Kit Components:
1. Akt Phosphorylated (Ser473) 1x40ul MW: 60kD
2. Akt Phosphorylated (Thr308) 1x40ul MW: 60kD
3. Akt, Pan 1x40ul MW: 60kD
4. c-Raf Phosphorylated (Ser259) 1x40ul MW: 74kD
5. GSK-3b Phosphorylated (Ser9) 1x40ul MW: 46kD
6. PTEN Phosphorylated (Ser380) 1x40ul MW: 54kD
7. PDK1 Phosphorylated (Ser241) 1x40ul MW: 58-68kD
8. LY294002 (PI3 Kinase Inhibitor) 1x300ug
9. IgG (HRP) GtxRb 1x100ul

Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Purity: Purified by protein A and peptide affinity chromatography.
Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, BSA, 50% glycerol.

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
..Franke, T.F. (1997) Cell 88, 435-437. (2) Burgering, B.T. and Coffer, P.J. (1995) Nature 376, 599-602. 3. Franke, T.F. et al. (1995) Cell 81, 727-736. 4. Alessi, D.R. et al. (1996) EMBO J. 15, 6541-6551. 5. Cardone, M.H. et al. (1998) Science 282, 1318-1321. 6. Brunet, A. et al. (1999) Cell 96, 857-868. 7. Cantley, L.C. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 4240-4245. 8. Vlahos, C. et al. (1994) J. Biol. Chem. 269, 5241-5248.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.