The serine/threonine kinase Akt family contains several members, including Akt1 (also designated PKBa or RacPK), Akt2 (also designated PKB-b or RacPK-b) and Akt3 (also designated PKB-g or thyoma viral proto-oncogene 3). Akt1 and Akt2 are activated by PDGF stimulation. This activation is dependent on PDGFR-b tyrosine residues 740 and 751, which bind the 85 kDa subunit of the phosphatidylinositol 3-kinase (PI 3-kinase) complex. Activation of Akt1 by insulin or insulin-growth factor-1 (IGF-1) results in phosphorylation of both Thr 308 and Ser 473. Phosphorylation of both residues is important to generate a high level of Akt1 activity, and the phosphorylation of Thr 308 is not dependent on phosphorylation of Ser 473 in vivo. Thus, Akt proteins become phosphorylated and activated in insulin/IGF-1- stimulated cells by an upstream kinase(s). The activation of Akt1 and Akt2 is inhibited by the PI kinase inhibitor wortmannin. Taken together, this data strongly suggests that the protein signals downstream of the PI kinases.
Applications:
Suitable for use in Western Blot.
Other applications not tested.
Recommended Dilution:
Western Blot: 1:1000
Optimal dilutions to be determined by the researcher.
Positive Control:
Untreated and PDGF treated NIH-3T3 fibroblasts.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees 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.
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