Western blot analysis of extracts from HEK293 cells untreated and treated with calyculin A (50 nM for 30 minutes), using Phospho-β-Catenin (upper) and β-Catenin Antibody #9562 (lower).
Western blot analysis of extracts from HEK293 cells untreated and treated with 50 nM calyculin A for 30 minutes, using Phospho-β-Catenin (Ser45) Antibody (lanes 1-2) or the same antibody after preincubation with listed peptides (lanes 3-8). (S45-P is phospho-Ser45 peptide; S33/37/T41-P is phospho-Ser33/37/Thr41 peptide; S45-non-P is nonphospho-Ser45 peptide.)
Western blot analysis of extracts from SW480 cells, using Phospho-β-Catenin (Ser45) Antibody.
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
Phospho-β-Catenin (Ser45) Antibody detects endogenous levels of beta-catenin only when phosphorylated at serine 45. It does not recognize β-catenin phosphorylated at other sites.
Mouse, Rat, Dog, Pig
Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser45 of human β-catenin. Antibodies are purified by protein A and peptide affinity chromatography.
β-Catenin is a key downstream effector in the Wnt signaling pathway (1). It is implicated in two major biological processes in vertebrates: early embryonic development (2) and tumorigenesis (3). CK1 phosphorylates β-catenin at Ser45. This phosphorylation event primes β-catenin for subsequent phosphorylation by GSK-3β (4-6). GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37, and Thr41 (7). Mutations at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines (8).
Explore pathways + proteins related to this product.