Technical Data
A1125-12A
Akt, phosphorylated (Ser505) (PKB, Rac)
Description:
Akt, also referred to as PKB or Rac, plays a critical role in controlling the balance between survival and apoptosis (13). 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 carboxy-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). One of the essential functions of Akt is the regulation of glycogen synthesis through phosphorylation and inactivation of glycogen synthase kinase-3alpha and beta (9,10). Akt may also play a role in insulin stimulation of glucose transport (9). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK3beta-mediated phosphorylation and degredation of cyclin D1 (11), and by negatively regulating the cyclin-dependent kinase inhibitors p27 KIP (12) and p21 WAF1 (13). Akt also plays a critical role in cell growth by directly phosphorylating the mammalian target of rapamycin, mTOR (14), but more importantly through phosophorylation and inactivation of Tuberin (TSC2), an mTOR inhibitor (15). Drosophila Akt (D-Akt) regulates multiple biological
processes such as cell survival (16) and cell growth (17). Major sites of phosphorylation include Ser505 and Thr342. These activation sites are homologous to mammalian
Ser473 and Thr308 respectively. Identified downstream targets of D-Akt include Trh (Ser665) (18) and Tsc2 (19). Like Akt, D-Akt functions in a wortmannin- sensitive pathway involving PI3 kinase (20).

Applications:
Suitable for use in Western Blotting, Immunocytochemistry. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000
Immunocytochemistry (IF) 1:500

Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and add glycerol (30-50%). Freeze at -20C. Aliquots are stable for at least 12 months at -20C. 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
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceDrosophilaRabbit
Concentration:
Not determined
Immunogen:
Synthetic phospho peptide (KLH coupled) corresponding to residues around ser505 of drosophila Akt
Purity:
Protein A and peptide affinity chromatography
Form
Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol
Specificity:
Recognizes endogenous levels of Akt only when phosphorylated at Ser505. Does not recognize drosophila Akt when phosphorylated at other sites, not does it recognize related kinases such as PKC or p70 S6 Kinase
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
(1) Franke, T.F. (1997) Cell 88, 435437. (2) Burgering, B.T. and Coffer, P.J. (1995) Nature 376, 599602. (3) Franke, T.F. et al. (1995) Cell 81, 727736. (4) Alessi, D.R. et al. (1996) EMBO J. 15, 65416551. (5) Cardone, M.H. et al. (1998) Science 282, 13181321. (6) Brunet, A. et al. (1999) Cell 96, 857868. (7) Cantley, L.C. et al. (1999) Proc. Natl. Acad. Sci. USA 96, 42404245. (8) Vlahos, C. et al. (1994) J. Biol. Chem. 269, 52415248. (9) Hajduch, E. et al. (2000) FEBS Lett. 492, 199203.(10) Cross, D.A. et al. (1995) Nature 373, 785789. (11) Diehl, J.A. et al. (1998) Genes Dev. 12, 34993511.