Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® GSK-3α siRNA II (+) or SignalSilence® GSK-3α siRNA I #6312 (+), using GSK-3α Antibody #9338 and β-Actin (13E5) Rabbit mAb #4970. GSK-3α Antibody confirms silencing of GSK-3α expression and β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of GSK-3α siRNA.
Western blot analysis of recombinant GST-GSK-3beta protein (76kD), and extracts from HeLa and PC3 cells, using GSK-3alpha Antibody (left) and GSK-3beta Antibody (right).
|REACTIVITY||H M R Mk|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
GSK-3alpha Antibody detects endogenous levels of total GSK-3alpha protein. It does not cross-react with recombinant GSK-3beta.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human GSK-3alpha. Antibodies are purified by protein A and peptide affinity chromatography.
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-3alpha regulates the production of amyloid-beta peptides, a major component of the plaques that accumulate with progression of Alzheimer's disease. Administration of therapeutic concentrations of lithium, a GSK-3 inhibitor, attenuates amyloid-beta production by specifically inhibiting the cleavage of amyloid precursor protein (APP) by gamma secretase, blocking accumulation of amyloid-beta peptides in the brains of mice that overproduce APP (6).
Explore pathways + proteins related to this product.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.