Product Pathways - Akt Signaling
Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb #9323
| Applications | Reactivity | MW (kDa) | Source | Isotype |
|---|---|---|---|---|
| W IHC-P IF-IC | H M (R) (Mk) | 46 | Rabbit | IgG |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb detects endogenous levels of GSK-3β only when phosphorylated at Ser9. The antibody may cross-react weakly with the phosphorylated form of GSK-3α due to high sequence homology.
Source / Purification
Monoclonal antibody is produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues around Ser9 of human GSK-3β.
Western Blotting
Western blot analysis of extracts from NIH/3T3 cells, λ-phosphatase- or PDGF-treated, using Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb (upper) or GSK-3β (27C10) Rabbit mAb #9315 (lower).
Western Blotting
Western blot analysis of extracts from wild type (lanes 1,2), GSK-3α knock out (lanes 3,4) and GSK-3β knock out (lanes 5,6) mouse embryonic fibroblast cells (MEF), untreated or PDGF treated, using Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb (upper) and GSK-3α/β Antibody (lower). (MEF wild type, GSK-3α knock out and GSK-3β knock out cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-GSK3β (Ser9) (5B3) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis using Phospho-GSK-3beta (Ser9) (5B3) Rabbit mAb on SignalSlide(TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded wild type (left) or GSK-3 beta knock out (right) MEF cell pellets using Phospho-GSK-3beta (Ser9) (5B3) Rabbit mAb. (MEF wild type, GSK-3alpha knock out and GSK-3beta knock out cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
IF-IC
Confocal immunofluorescent analysis of wild type (left), GSK-3alpha knock out (center), or GSK-3beta knock out (right) MEF cells, FGF-treated (top) or phosphatase-treated (bottom) using Phospho-GSK-3beta (Ser9) (5B3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye). (MEF wild type, GSK-3alpha knock out and GSK-3beta knock out cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).
Background
Glycogen synthase kinase-3 (GSK-3) was initially identified as an enzyme that regulated 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 PI3 kinase/Akt cell survival pathway, and its 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.
Application References
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Companion Products
- 9316 Phospho-GSK-3α (Ser21) (36E9) Rabbit mAb
- 9327 Phospho-GSK-3α/β (Ser21/9) (37F11) Rabbit mAb (GSK-3α Preferred)
- 9338 GSK-3alpha Antibody
- 9315 GSK-3β (27C10) Rabbit mAb
- 9270 PhosphoPlus® Akt (Ser473) Antibody Kit
- 9916 Phospho-Akt Pathway Sampler Kit
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8112 SignalStain® Antibody Diluent
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.
