Titlebook: Wnt Signaling; Methods and Protocol Quinn Barrett,Lawrence Lum Book 2016 Springer Science+Business Media New York 2016 Regenerative medicin

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Visualizing Wnt Palmitoylation in Single Cells,gating Wnt activity, secretion, and its interaction with cellular membrane compartments. This protocol describes a method we have recently developed to detect cellular Wnt palmitoylation. The method, combining click chemistry, bio-orthogonal fatty acid probes, and proximity ligation assay (PLA), pro

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Monitoring Wnt Protein Acylation Using an In Vitro Cyclo-Addition Reaction,S-PAGE. This protocol incorporates in vivo labeling of a Wnt-IgG Fc fusion protein using an alkynylated palmitate probe but departs from a traditional approach by incorporating a secondary cycloaddition reaction performed on single-step purified Wnt protein immobilized on protein A resin. This appro

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Methods for Studying Wnt Protein Modifications/Inactivations by Extracellular Enzymes, Tiki and Nototeases to cleave a fragment from the amino terminus of Wnt proteins. Notum is a Wnt deacylase that removes the lipid modification that is essential for Wnt activities. Here, we provide detailed procedures for preparing enzymatic active Tiki and Notum proteins and the in vitro enzymatic reactions. W

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Probing Wnt Receptor Turnover: A Critical Regulatory Point of Wnt Pathway,trol proper signaling output. Recent studies suggest that cell surface Wnt receptor level is controlled by ubiquitination, and serve as a critical regulatory point of Wnt pathway activity as it determines the responsiveness of cells to Wnt signal. Here, we describe flow cytometry, cell surface prote

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The Use of Chick Embryos to Study Wnt Activity Gradients,troporation can be directed to a single side of the spinal cord, thus leaving the contralateral side as an internal control. Here, we describe a method for measuring Wnt signaling via the use of BAT-Gal, a β-catenin dependent Wnt reporter.

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Monitoring Wnt Signaling in Zebrafish Using Fluorescent Biosensors,Our technology is based on artificial enhancers, obtained by polymerization of TCF binding elements, cloned upstream to ubiquitous or tissue-specific promoters. The different promoter/enhancer combinations are used to drive fluorescent protein reporter constructs integrated in the zebrafish germline

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