Wnt3a (Wingless-type MMTV (Mouse Mammary Tumor Virus) Integration Site Family Member)

The Wnt family of consists of ligands involved in cell development, fate determination, organogenesis and oncogenesis. Information on Wnt signaling has been inferred from genetic analysis of wingless (wg) signaling in Drosophila. The Wnt signaling cascade initiates at the cell membrane when Wnt interacts with its receptor, a member of the Frizzled (Fz) transmembrane receptor family and lipoprotein receptor-related proteins 5 and 6 (LRP-5/6). This interaction relays the signal into the interior of the cell where disheveled (dsh) is brought to the membrane and gets phosphorylated/activated. Dsh promotes the inactivation of GSK3 kinase activity, thus preventing the phosphorylation of beta-Catenin and allowing it to accumulate as it avoids ubiquination and subsequent proteosome degradation. Stable beta-Catenin accumulates in the cytoplasm where it interacts with lymphoid enhancer factor/T cell factor (LEF/TCF) and translocates into the nucleus. This results in activation of TCF-responsive genes which are thought to play key roles in development and cancer. Negative regulators of the pathway may include axin and the tumor suppressor APC. Applications: Suitable for use in Immunohistochemistry and Western Blot. Other applications not tested. Recommended Dilutions: Immunohistochemistry (Paraffin): 1:200 stains whole-mount transverse section of E12.5 mouse embryos. Incubated with diluted W3011-02B for 1 hour at RT. Tissue was pretreated with no heat induced epitope retrieval, citrate pH 6. Western Blot: 1:1000. Rat whole brain lysate was resolved by electrophoresis and transferred to PVDF. Proteins were visualized using an IgG (HRP) Dk xRb and a chemiluminescence detection system. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.