|Detection of Human and Mouse Wnt‑5a by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and mouse embryo (13 d.p.c.) tissue. PVDF Membrane was probed with 2 µg/mL of Human/Mouse Wnt‑5a Monoclonal Antibody (Catalog # MAB645) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Wnt‑5a at approximately 49 kDa (as indicated). This experiment was conducted under non-reducing conditions and using Immunoblot Buffer Group 1.|
|Wnt‑5a in Mouse Embryo. Wnt‑5a was detected in immersion fixed frozen sections of mouse embryo using Human/Mouse Wnt‑5a Monoclonal Antibody (Catalog # MAB645) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti‑Rat IgG Secondary Antibody (orange; Catalog # NL013) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.|
Wnt proteins are secreted glycoproteins that contain a conserved pattern of 23‑24 cysteine residues. Wnts play critical roles in both carcinogenesis and embryonic development for a variety of organisms. Wnts bind to receptors of the Frizzled family, sometimes in conjunction with other membrane-associated proteins such as LRPs or proteoglycans. Downstream effects of Wnt signaling occur through different intracellular components, depending on which pathway is activated. Three pathways have been characterized: the canonical Wnt/ beta -catenin pathway, the Wnt/Ca2+ pathway, and the planar cell polarity (1‑2).
Wnt-5a is part of the subgroup of Wnts that are not axis-inducing in Xenopus embryos and do not transform C57MG mammary epithelial cells. This subgroup is also implicated in the Wnt/Ca2+ pathway, playing roles in cell movements and cell adhesion (3). This non-canonical Wnt pathway can inhibit canonical Wnt/ beta -catenin signaling. In Wnt-5a deficient mouse embryos, beta -catenin accumulates in the limb bud suggesting that Wnt-5a normally promotes degradation of beta -catenin (4). Likewise, in Xenopus embryos Wnt-5a antagonizes the ability of the canonical Wnt subgroup to induce a secondary axis (5). Wnt-5a is implicated in various types of cancer and has complex roles. It acts as a tumor suppressor for mammary, B-cell, colon, and uroepithelial cancer cells but is up-regulated in melanomas, where expression levels correlate with severity of metastasis (3). Furthermore, aberrant Wnt-5a signaling results in other diseases such as rheumatoid arthritis (6). Like other developmental growth factors Wnt-5a has diverse roles in development. They are too numerous to enunciate here, as functions span from early anterior-posterior development and gastrulation movements to maintaining hematopoietic stem cell population, lung morphogenesis, and limb outgrowth. Mature Wnt-5a is a 49 kDa protein that shares 99% amino acid identity in mouse, rat and human.
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