Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with TPA #4174 (200 nM, 30 min; +), using Phospho-BCL9L (Ser915) Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (100 μM, 2 hr; +), using Phospho-BCL9L (Ser915) Antibody (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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-BCL9L (Ser915) Antibody recognizes endogenous levels of BCL9L protein only when phosphorylated at Ser915.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser915 of human BCL9L protein. Antibodies are purified by protein A and peptide affinity chromatography.
B-cell CLL/lymphoma 9-like protein (BCL9L, BL2, Bcl9-2, DLNB11) is a transcriptional activator that was originally identified in silico based on homology to BCL9 (1). BCL9L was subsequently found to play an important role in Wnt/β-catenin signaling by interacting with β-catenin and enhancing the transactivation potential of the β-catenin/TCF complex (2). Research studies show that BCL9L can increase the tumorigenic effect of aberrant Wnt signaling in some cases of colorectal cancer (2). Expression of BCL9L is correlated with tumor progression in colorectal (3) and breast cancer (4). Targeted deletion of BCL9 and BCL9L in the intestinal epithelium resulted in abrogation of Wnt target genes, including those controlling epithelial-mesenchymal transition and stem-cell like properties (5).
Phospho-BCL9L (Ser915) Antibody is directed at a site that was identified at Cell Signaling Technology (CST) using PhosphoScan®, CST's LC-MS/MS platform for modification site discovery. Phosphorylation at Ser915 was discovered using an AMPK substrate antibody. Please visit PhosphoSitePlus®, CST's modification site knowledgebase, at www.phosphosite.org for more information.
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