Product Pathways - PI3K / Akt Signaling
Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb #5558
|W IP IF-IC F||H M R Hm||Endogenous||46||Rabbit IgG|
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb detects endogenous levels of GSK-3β only when phosphorylated at Ser9. This antibody reacts with denatured components of bovine serum, including BSA.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser9 of human GSK-3β.
Western blot analysis of extracts from GSK-3α (-/-) (lanes 1,2) and GSK-3β (-/-) (lanes 3,4) mouse embryonic fibroblast (MEF) cells, λ phosphatase or PDGF-treated, using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (upper) and α/β-Tubulin Antibody #2148 (lower). (MEF wild type, GSK-3α (-/-) and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (upper) or GSK-3β (27C10) Rabbit mAb #9315 (lower).
Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or PDGF-treated (green), using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb.
Confocal immunofluorescent analysis of wild type mouse embryonic fibroblasts (MEFs) (top row), GSK-3β (-/-) MEFs (middle row) , or PC-3 cells (bottom row), untreated (left), LY294002- and Wortmannin-treated (#9901 and #9951 respectively; center) or lambda phosphatase-treated (right), using Phospho-GSK-3β (Ser9) (D85E12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild type and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
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.
- Mirlashari, M.R. et al. (2012) Leuk Res 36, 499-508. Applications: Flow Cytometry
- Biswas, K. et al. (2013) J Biol Chem 288, 2325-39. Applications: Western Blotting
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- 7003 20X LumiGLO® Reagent and 20X Peroxide
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- 9998 BSA
- 9999 Nonfat Dry Milk
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For Research Use Only. Not For Use In Diagnostic Procedures.