Titlebook: Notch Signaling; Methods and Protocol Hugo J. Bellen,Shinya Yamamoto Book 2014 Springer Science+Business Media New York 2014 Notch activity

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Tracking Trafficking of Notch and Its Ligands in Mammalian Cells,n Notch signaling activation and modulation. In mammalian cultured cells it is possible to track these cell surface molecules by pulse-labeling these proteins in vivo. One labeling protocol consists in the covalent linkage of membrane-impermeable biotin followed by western blotting. An alternative p

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Visualizing Notch Signaling In Vivo in , Tissues, have been developed to enable monitoring of pathway activity in ., including endogenous Notch-responsive genes and synthetic transcriptional reporter constructs. Here we summarize some of the different Notch signaling reporters that are available, discuss their relative merits, and describe two met

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Monitoring Notch Activity in the Mouse,ifying Notch signaling with high temporal and spatial resolution. This research has contributed substantially to our understanding of the role of Notch in cell specification and cellular physiology. Here, we present two protocols to monitor Notch activity in the mouse brain: (1) by intraventricular

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Notch Signaling Assays in , Cultured Cell Lines,sessing whether specific enhancers or genes are regulated by Notch. In this chapter, we will describe two different types of assays that can be used to monitor Notch activation in Kc167 and S2 cells. One involves activating Notch in cultured cells and measuring the change in endogenous gene expressi

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Monitoring Notch Activation in Cultured Mammalian Cells: Transcriptional Reporter Assays,rive reporter gene expression provides a flexible and robust approach for monitoring signaling activity. In this chapter, we present an overview of Notch transcriptional reporter assays and discuss different methods for ligand-independent and ligand-dependent activation of Notch in mammalian cells.

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Monitoring Notch Activation in Cultured Mammalian Cells: Luciferase Complementation Imaging Assays,ing partner CSL to recruit the coactivator MAML and additional cofactors to ultimately activate target gene expression. Taking advantage of the specific interaction between NICD and these factors, we have developed a luciferase complementation imaging (LCI)-based reporter system to quantitatively mo

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Proteomic Analysis of the Notch Interactome,r several model organisms. Defining protein interactions on a proteome-wide scale has led to a better understanding of the cellular functions of many proteins, especially those that have not been studied by classical molecular genetic approaches. Here we describe the resources, methods, and techniqu

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