Notch Signaling: Methods and Protocols focuses on molecular, biochemical and bioinformatics aspects of Notch signaling. Chapters analyze the Notch interactome, post-translational modifications of Notch, ligand binding assays, methods to assess proteolytic cleavage and transcriptional targets. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Notch Signaling: Methods and Protocols is a valuable contribution to hundreds of labs and thousands of scientists who pursue this research area with vigor.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt
Introduction to Notch Signaling.- Genetic Screens to Identify New Notch Pathway Mutants in Drosophila.- Structure-function Analysis of Drosophila Notch using Genomic Rescue Transgenes.- Overview of Genetic Tools and Techniques to Study Notch Signaling in Mice.- Immunohistochemical Tools and Techniques to Visualize Notch in Drosophila melanogaster.- Antibody Uptake Assay and in vivo Imaging to Study Intracellular Trafficking of Notch and Delta in Drosophila.- Tracking Trafficking of Notch and its Ligands in Mammalian Cells.- Visualizing Notch Signaling in vivo in Drosophila Tissues.- Monitoring Notch Activity in the Mouse.- Notch Signaling Assays in Drosophila Cultured Cell Lines.- Monitoring Notch Activation in Cultured Mammalian Cells: Transcriptional Reporter Assays.- Monitoring Notch Activation in Cultured Mammalian Cells: Luciferase Complementation Imaging Assays.- Visualization of Notch Signaling Oscillation in Cells and Tissues.- Proteomic Analysis of the Notch Interactome.- Bacterial Expression and in vitro Refolding of Limited Fragments of the Notch Receptor and its Ligands.- Analyzing the Post-translational Modification Status of Notch using Mass Spectrometry.- Assay to Probe Proteolytic Processing of Notch by -secretase.- Analyzing the Nuclear Complexes of Notch Signaling by Electrophoretic Mobility Shift Assay.- Identifying Direct Notch Transcriptional Targets using the GSI-washout Assay.- Probing the Epigenetic Status at Notch Target Genes.- Notch-ligand Binding Assays in Drosophila Cells.- Modeling Notch Signaling: A Practical Tutorial.- Small Molecules that Inhibit Notch Signaling.- Application and Evaluation of Anti-Notch Antibodies to Modulate Notch Signaling.- Application of Anti-ligand Antibodies to Inhibit Notch Signaling.