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You are here:Home » Molecular Biology » MB-Protein Kinases » Akt, phosphorylated (Thr308) Blocking Peptide

Akt, phosphorylated (Thr308) Blocking Peptide

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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 by phosphorylation within the carboxy-terminus at Ser473. AKT promotes cell survival by inhibiting apoptosis through its ability to phosphorylate and inactivate several targets, including
Bad (5), Forkhead transcription factors (6), c-Raf (7) and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/AKT signaling pathway (8).
Catalog #A1124-92A
LY294002 is a specific PI3 kinase inhibitor (9). One of the essential functions of AKT is the regulation of glycogen synthesis through phosphorylation and inactivation of glycogen synthase kinase-3a and b 10,11). AKT may also play a role in insulin stimulation of glucose transport (10). In addition to its role in survival and glycogen synthesis, AKT is involved in cell cycle regulation by preventing GSK3b-mediated phosphorylation and degradation of cyclin D1 (12), and by negatively regulating the cyclin-dependent kinase inhibitors p27 KIP (13) and p21 WAF1 (14). AKT also plays a critical role in cell growth by directly phosphorylating the mammalian target of rapamycin, mTOR (15), but more importantly through phosphorylation and inactivation of tuberin TSC2), an mTOR inhibitor (16). 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 (4EEP1), an inhibitor of translation (17,18)
Applications Applications: Use as a blocking reagent to evaluate the specificity of antibody reactivity in Western immunoblotting and immunohistochemistry protocols.
ProtocolWestern blotting: Add 10ul of antibody and 10ul of blocking peptide to 10ml of antibody dilution buffer, and incubate at room temperature for 2 hours before allowing to react with the blot.
Immunohistochemistry Add 2ul of antibody and 5ul of blocking peptide to 153ul of buffer, and
incubate at least 30 minutes before allowing to react with the slides.
Storage and StabilityMay be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 6 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.
PurityPurified
Concentration100ug
(10 Western mini-blots)
FormSupplied as a liquid in 20 mM potassium phosphate, pH 7.0, 50 mM NaCl, 0.1 mM EDTA, 1mg/ml BSA and 5% glycerol.
SpecificityPeptide specificity: Human, mouse
Important NoteThis 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.


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