Product Pathways - PI3K / Akt Signaling
GSK-3 Fusion Protein #9237
GSK-3 Fusion Protein serves as a useful substrate for assaying Akt kinase activity. It is expressed as a protein fusion to the GSK-3 α/β crosstide, corresponding to residues surrounding GSK-3 α/β (Ser21/9) (CGPKGPGRRGRRRTSSFAEG).
AKT Kinase activity of PDGF-treated NIH/3T3 cell extracts was analyzed by IP/Kinase assay. Cell extracts (200 μl) were incubated overnight with Immobilized Phospho-Akt (Ser473) (D9E) Rabbit mAb #4070. After extensive washing the kinase reaction was performed in the presence of 200 μM of cold ATP and 1 μg of GSK-substrate. Phosphorylation of GSK-3 was measured by Western blot using Phospho-GSK-3 a/β (Ser21/9) Antibody #9327.
Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulates glycogen synthesis in response to insulin (1). GSK-3 is a ubiquitously expressed serine/threonine protein kinase that phosphorylates and inactivates glycogen synthase. GSK-3 is a critical downstream element of the PI3K/Akt cell survival pathway whose activity can be inhibited by Akt-mediated phosphorylation at Ser21 of GSK-3α and Ser9 of GSK-3β (2,3). GSK-3 has been implicated in the regulation of cell fate in Dictyostelium and is a component of the Wnt signaling pathway required for Drosophila, Xenopus, and mammalian development (4). GSK-3 has been shown to regulate cyclin D1 proteolysis and subcellular localization (5).
- Welsh, G.I. et al. (1996) Trends Cell. Biol. 6, 274-279.
- Srivastava, A.K. and Pandey, S.K. (1998) Mol. Cell. Biochem. 182, 135-141.
- Cross, D.A. et al. (1995) Nature 378, 785-789.
- Nusse, R. (1997) Cell 89, 321-323.
- Diehl, J.A. et al. (1998) Genes Dev. 12, 3499-3511.
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- 9840 Akt Kinase Assay Kit (Nonradioactive)
- 9336 Phospho-GSK-3β (Ser9) Antibody
- 9327 Phospho-GSK-3α/β (Ser21/9) (37F11) Rabbit mAb (GSK-3α Preferred)
For Research Use Only. Not For Use In Diagnostic Procedures.