Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - PI3K / Akt Signaling

GSK-3α/β (D75D3) XP® Rabbit mAb #5676

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H M R Hm Mk Endogenous 51, 46 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  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

GSK-3α/β (D75D3) XP® Rabbit mAb detects endogenous levels of total GSK-3α and GSK-3β. The peptide sequence used as the antigen is 100% conserved between GSK-3α and GSK-3β in humans, monkeys, mice and rats. The antibody may prefer the conformation of GSK-3β by IF-IC.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Gln269 of human GSK-3α protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with SignalSilence® GSK-3α/β siRNA #6301, using GSK-3α/β (D75D3) XP® Rabbit mAb (upper) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from GSK-3β (-/-) (lanes 1,2), GSK-3α (-/-) (lanes 3,4) and wild type (lanes 5,6) mouse embryonic fibroblast cells (MEF), untreated or insulin treated, using GSK-3α/β (D75D3) XP® Rabbit mAb. (MEF wild type, GSK-3α (-/-) and GSK-3β (-/-) cells were kindly provided by Dr. Jim Woodgett, University of Toronto, Canada).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, mock-transfected (left) or transfected with SignalSilence® GSK-3α/β siRNA #6301 (right), using GSK-3α/β (D75D3) XP® Rabbit mAb (green). Actin filaments have been labeled wth DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


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).

GSK-3α regulates the production of amyloid-β peptides, a major component of the plaques that accumulate with progression of Alzheimer disease. Administration of therapeutic concentrations of lithium, a GSK-3 inhibitor, attenuates amyloid-β production by specifically inhibiting the cleavage of amyloid precursor protein (APP) by γ-secretase, blocking accumulation of amyloid-β peptides in the brains of mice that overproduce APP (6).

  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.
  6. Phiel, C.J. et al. (2003) Nature 423, 435-9.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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