antibody

Akt (Rac PKa, PKBa) (pan)

A1124-03F

100 ul

monoclonal

domestic rabbit

nonconjugated

8A4(C67E7)

Akt (Rac PKa, PKBa) (pan)

rabbit

Mab

Antibodies

100 ul

8A4(C67E7)

IgG

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 by phosphorylating and inactivating 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 throughp phosphorylation and inactivation of GSK-3alpha and beta (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-3beta mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip (15) and p21 Waf1 (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). Inhibition of mTOR stops the protein synthesis machinery due to inactivation of its effector, p70 S6 kinase and activation of the eukaryotic initiation factor, 4E binding protein 1 (4E-EP1), an inhibitor of translation (19,20). Applications: Suitable for use in Immunofluorescence, Flow Cytometry, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested. Recommended Dilution: Immunofluorescence (IF-IC): 1:400 Flow Cytometry: 1:100 Western Blot: 1:1000 Immunoprecipitation: 1:50 Immunohistochemistry (Paraffin): 1:300 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Hu Mk Mo Rt

Synthetic peptide corresponding to the C-terminal sequence of mouse Akt (KLH).

Recognizes mouse Akt. Detects endogenous levels of total Akt protein. Does not crossreact with other related proteins. Species Crossreactivity: Human, monkey and rat.

Serum

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol, 0.02% sodium azide.

Not determined

Product Type: Mab Isotype: IgG Clone No: 8A4(C67E7) Host: rabbit Source: mouse Concentration: Not determined Form: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol, 0.02% sodium azide. Purity: Serum Immunogen: Synthetic peptide corresponding to the C-terminal sequence of mouse Akt (KLH). Specificity: Recognizes mouse Akt. Detects endogenous levels of total Akt protein. Does not crossreact with other related proteins. Species Crossreactivity: Human, monkey and rat. 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.

FC IF IHC IP WB

-20°C

1. Franke, T.F., et al., Cell 88: 435–437 (1997). 2. Burgering, B.T. and Coffer, P.J., Nature 376: 599–602 (1995). 3. Franke, T.F., et al., Cell 81: 727–736 (1995). 4. Alessi, D.R., et al., EMBO J. 15: 6541–6551 (1996). 5. Sarbassov, D.D., et al., Science 307: 1098–1101 (2005). 6. Jacinto, E., et al., Cell 127: 125–137 (2006). 7. Cardone, M.H., et al., Science 282: 1318–1321 (1998). 8. Brunet, A., et al., Cell 96: 857–868 (1999). 9. Zimmerman, S. and Moelling, K., Science 286: 1741–1744 (1999). 10. Cantley, L.C. and Neel, B.G., Proc. Natl. Acad. Sci. USA 96: 4240–4245 (1999). 11. Vlahos, C., et al., J. Biol. Chem. 269: 5241–5248 (1994). 12. Hajduch, E., et al., FEBS Lett. 492: 199–203 (2000). 13. Cross, D.A., et al., Nature 373: 785–789 (1995). 14. Diehl, J.A., et al., Genes Dev. 12: 3499–3511 (1998). 15. Gesbert, F., et al., J. Biol. Chem. 275: 39,223–39,230 (2000). 16. Zhou, B.P., et al., Nat. Cell Biol. 3: 245–252 (2001). 17. Nave, B.T., et al., Biochem. J. 344: 427–431 (1999). 18. Manning, B.D., et al., Mol. Cell 4: 648–657 (2000). 19. Manning, B.D., et al., Mol. Cell 10: 151–162 (2002). 20. Inoki, K., et al., Nat. Cell Biol. 4: 648–657 (2002).

https://usbio-images.r.worldssl.net/prodimages/14/A1124-03F_1.jpg

Confocal immunofluorescent analysis of C2C12 cells, LY294002- treated (left) or insulin-treated (right), using A1124-03F Akt (pan) Rabbit mAb (green).