Cell Signaling Technology

Product Pathways - PI3K / Akt Signaling

Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb #9323

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IF-IC H M R Mk Endogenous 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 cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

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 animals with a synthetic phosphopeptide corresponding to residues around Ser9 of human GSK-3β.

Western Blotting

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 Blotting

Western blot analysis of extracts from wild type (lanes 1,2), GSK-3α (-/-) (lanes 3,4) and GSK-3β (-/-) (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 (-/-) and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-GSK3β (Ser9) (5B3) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis using Phospho-GSK-3β (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)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded wild type (left) or GSK-3β (-/-) MEFs, using Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb. (MEF wild type, GSK-3α knock out and GSK-3β knock out cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).

IF-IC

IF-IC

Confocal immunofluorescent analysis of wild type (left), GSK-3α (-/-) (center), or GSK-3β (-/-) (right) mouse embryonic fibroblasts (MEFs), FGF-treated (top) or phosphatase-treated (bottom), using Phospho-GSK-3β (Ser9) (5B3) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild type, GSK-3α (-/-) and GSK-3β (-/-) 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 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).

  1. Welsh, G.I. et al. (1996) Trends Cell. Biol. 6, 274-279.
  2. Srivastava, A.K. and Pandey, S.K. (1998) Mol. Cell. Biochem. 182, 135-141.
  3. Cross, D.A. et al. (1995) Nature 378, 785-789.
  4. Nusse, R. (1997) Cell 89, 321-323.
  5. Diehl, J.A. et al. (1998) Genes Dev. 12, 3499-3511.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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