Akt, phosphorylated (Thr-34)

Akt, also known as Protein Kinase B (PKB) and Rac (Related to the A and C kinases), is a 60kD serine/threonine kinase critical for controlling diverse cellular functions, including glucose metabolism, gene transcription, cell proliferation, and apoptosis. Akt phosphorylates a number of physiological substrates including MBP, glycogen synthetase, PKA RII subunit, and histone H1. Akt is activated in response to insulin and growth factors in a PI3-kinase dependent manner. Activation of PI3-Kinase generates phosphatidylinositol 3,4-bisphosphate which may induce the membrane translocation of Akt coincident with its phosphorylation and activation. Upon activation, Akt associates with members of the PKC family of kinases, such as PKCd and PKCz. Ceramide-activated PKCz leads to phosphorylation of the Akt-Pleckstrin Homology (PH) domain on threonine 34. This phosphorylation inhibits PIP3 binding to Akt, preventing activation of the kinase by insulin. Since Akt confers a prosurvival signal and regulates pathways in response to insulin, suppressing its activation may be one mechanism by which ceramide promotes cell death and induces insulin resistance.\n\nApplications: \nSuitable for use in ELISA and Western Blot. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 1:1000. Membrane was incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween-20 for 1 hour at room temperature.\nELISA: 1:2000.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees 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.\n

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SPECIFICATIONS

Catalog Number

A1124-09

Size

100ul

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Hum, Rat, Rabbit

Form

Supplied as a liquid in PBS, 50% glycerol, 1mg/ml BSA, and 0.05% sodium azide.

P Type

Pab

Purity

Purified by affinity chromatography.

References

1. 1991 Jones, P.F. et al. Proc. Natl. Acad. Sci. USA 88:4171-4175. 2. 1997 Marte, B. & Downward. J. TIBS. 22:355-358. 3. 2003 Powell, D.J. et al. Mol. Cell Biol. 23:7794-7808.

Additional Info

Recognizes a 60kD protein corresponding to the apparent molecular mass of phosphorylated Akt on SDS-PAGE immunoblots of A431+ calyculin A cell lysate. Similar results were seen in calyculin A treated human aortic endothelial and HeLa cells, rabbit spleen fibroblasts, and rat pituitary cells.