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-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/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).
Inhibition of mTOR stops the protein synthesis machinery by inactivating p70 S6 kinase and activating the eukaryotic initiation factor 4E binding protein 1 (4E-BP1), an inhibitor of translation (18,19).
JNK reactivates Akt after ischemic injury by phosphorylating Thr450, a priming event for subsequent phosphorylation by 3-phosphoinositide-dependent protein kinase (20).
Applications:Suitable for use in Western Blot and Immunoprecipitation.
Other applications not tested.
Recommended Dilution:Western Blot: 1:1000Immunoprecipitation: 1:50Optimal 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.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A and peptide affinity chromatography.
Form：Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 0.1mg/ml BSA, 50% glycerol.
Specificity：Recognizes endogenous levels of human Akt1 only when phosphorylated at Thr450. This antibody also recognizes Akt2 and Akt3 when phosphorylated at the corresponding residues. It does not recognize Akt phosphorylated at other sites. Species crossreactivity: Mouse and rat.
Isotype：IgG
Calc Applications Abbrev：IP WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：Synthetic phosphopeptide corresponding to residues surrounding Thr451 of Akt2.
