Product Pathways - Akt Signaling
SignalSilence® GSK-3α/β siRNA #6301
Western Blotting
Western blot analysis of extracts from HeLa cells, untransfected or transfected with GSK-3alpha/beta siRNA. GSK-3beta was detected using GSK-3beta Antibody #9332, and Bcl-xL was detected using Bcl-xL Antibody #2762. The GSK-3beta Antibody confirms silencing of GSK-3beta expression, and the Bcl-xL Antibody is used to control for loading and siRNA specificity.
Western Blotting
Western blot analysis of extracts from HeLa cells, untransfected or transfected with either GSK-3alpha/beta siRNA or GSK-3alpha siRNA. GSK-3alpha and GSK-3beta were detected using a GSK-3alpha/beta antibody, and eIF4B was detected using eIF4B Antibody #3592. The GSK-3alpha/beta antibody confirms silencing of GSK-3alpha and beta expression, and the eIF4B Antibody is used to control for loading and siRNA specificity.
Fluorescent Detection
Fluorescent detection of SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 in living HeLa cells 24 hours post-transfection, demonstrating nearly 100% transfection efficiency.
Directions for Use
CST recommends transfection with 50 nM GSK-3alpha/beta siRNA 48 hours prior to cell lysis. See Protocol for transfection procedure.
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 an 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).
Small Interfering RNA (siRNA) has been used to specifically silence GSK-3alpha and beta expression, causing an upregulation of beta-catenin protein levels (6).
- 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.
- Yu, J. Y. et al. (2003) Mol. Ther. 7, 228-236.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9331 Phospho-GSK-3α/β (Ser21/9) Antibody
- 9336 Phospho-GSK-3β (Ser9) Antibody
- 6300 SignalSilence® GSK-3α/β siRNA Kit
- 6201 SignalSilence® Control siRNA (Fluorescein Conjugate)
- 9315 GSK-3β (27C10) Rabbit mAb
Limited Use Label License, RNA interference: This product is licensed under European Patent 1144623 and foreign equivalents from Ribopharma AG, Kulmbach, Germany and is provided only for use in non-commercial research specifically excluding use (a) in drug discovery or drug development, including target identification or target validation, by or on behalf of a commercial entity, (b) for contract research or commercial screening services, (c) for the production or manufacture of siRNA-related products for sale, or (d) for the generation of commercial databases for sale to Third Parties. Information about licenses for these and other commercial uses is available from Ribopharma AG, Fritz-Hornschuch-Str. 9, D-95326 Kulmbach, Germany.
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
