|Detection of Human Wnt‑10b by Western Blot. Western blot shows lysates of SW480 human colorectal adenocarcinoma cell line and MDA‑MB‑468 human breast cancer cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Wnt‑10b Monoclonal Antibody (Catalog # MAB7196) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Wnt‑10b at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Detection of Human Wnt‑10b by Simple WesternTM. Simple Western lane view shows lysate of SW480 human colorectal adenocarcinoma cell line, loaded at 0.5 mg/mL. A specific band was detected for Wnt‑10b at approximately 58 kDa (as indicated) using 5 µg/mL of Mouse Anti-Human Wnt‑10b Monoclonal Antibody (Catalog # MAB7196). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.|
Wnt-10b (also known as Wnt-12) is a 42‑44 kDa member of the Wnt family of secreted, highly conserved, cysteine-rich glycoproteins that play important roles in vertebrate pattern formation, cell fate decision, axon guidance, and tumor formation (1‑3). Human Wnt-10b cDNA encodes a 389 amino acid (aa) precursor that contains a 28 aa signal sequence plus a 361 aa mature protein that contains two glycosylation sites, three potential phosphorylation sites, and a potential palmitoylation site (3, 4). Human Wnt-10b shares 97‑99% aa identity with mouse, rat, equine, porcine, and canine Wnt-10b. Wnt-10b plays a critical role in maintaining mesenchymal stem cells and determining whether they differentiate to adipocytes or osteoblasts (5‑7). Mouse Wnt-10b deletion produces age‑dependent loss of bone mass due to defective production of osteoblasts, while transgenic over‑expression increases postnatal osteoblast differentiation and inhibits adipocyte differentiation (5‑7). Ectopic expression of Wnt-10b in an obesity and diabetes‑prone background, such as the ob/ob mouse, inhibits obesity (8). In mouse skeletal muscle, Wnt-10b is expressed inversely with SREBP1c and increases insulin sensitivity (9). In humans, a missense polymorphism is responsible for a malformation of hands and feet, while a C256Y inactivating mutation is associated with severe early-onset obesity (10, 11). Wnt-10b is mainly produced by stem cells and pre-osteoblasts, but also by adult bone marrow CD8+ T lymphocytes stimulated with parathyroid hormone (12). In some hepatocellular carcinomas, Wnt-10b can inhibit cancer cell growth, but in others, it can act synergistically with FGFs to stimulate cell growth (13). Several Wnts, including Wnt-10b, are expressed in both normal and/or malignant colon tissues (14).