Western blot analysis of total cell lysates from HeLa, LN18, PANC1 cells and HeLa cells conditioned medium (CM) using Wnt5a/b (C27E8) Rabbit mAb.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. 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
Wnt5a/b (C27E8) Rabbit mAb detects endogenous levels of Wnt5a and Wnt5b proteins. This antibody cross-reacts with an unidentified protein between 55 and 60 kDa.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Glu201 of Isoform 1 and Glu186 of Isoform 2 of the human Wnt5a protein. This antibody works to detect both isoforms of human Wnt5a protein.
The Wnt family includes several secreted glycoproteins that play important roles in animal development (1). There are 19 Wnt genes in the human genome that encode functionally distinct Wnt proteins (2). Wnt members bind to the Frizzled family of seven-pass transmembrane proteins and activate several signaling pathways (3). The canonical Wnt/β-catenin pathway also requires a coreceptor from the low-density lipoprotein receptor family (4). Aberrant activation of Wnt signaling pathways is involved in several types of cancers (5).
Wnt-5a has been shown to signal through the canonical Wnt pathways as well as through non-canonical pathways and is up-regulated in various types of human cancers (6-8). In melanoma, Wnt5a is thought to directly affect cell motility and metastasis (9).
Explore pathways + proteins related to this product.