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This volume brings together cutting-edge laboratory protocols to characterize the novel fluorescent proteins (FPs) and approaches based on fluorescent proteins that aim to answer some of the key cell biological questions. The book covers topics ranging from the database of fluorescent proteins to their characterization and adaptation to a wide range of biological systems. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Fluorescent Proteins: Methods and Protocols serves as an ideal guide for students and academicians enthusiastic about the recent progress in the practical application of fluorescent protein technology.
List of contents
Using FPbase: The Fluorescent Protein Database.- Selecting the Fluorescent Protein for In Vivo Imaging Experiments.- Advanced Fluorescence Microscopy Methods to Study Dynamics of Fluorescent Proteins In Vivo.- Directed Evolution of Fluorescent Proteins in Bacteria.- Fluorescent Proteins: Crystallization, Structural Determination, and Non-Natural Amino Acid Incorporation.- Development and Characterization of Flavin-Binding Fluorescent Proteins, Part I: Basic Characterization.- Development and Characterization of Flavin-Binding Fluorescent Proteins, Part II: Advanced Characterization.- CRISPR-Cas9-Mediated Knock-In Approach to Insert the GFP11 Tag into the Genome of a Human Cell Line.- Fluorescent Protein-Based Approaches for Sub-Cellular Protein Localization in Plants.- Growth Rates Evaluation of the Budding Yeast Saccharomyces cerevisiae Cells Carrying Endogenously Expressed Fluorescent Protein Fusions.- DNA Visualization Using Fluorescent Proteins.- Quantification ofIntracellular Citrate Concentrations with Genetically Encoded Biosensors.- Quantification of Redox-Sensitive GFP Cysteine Redox State via Gel-Based Read-Out.- Assessing In Vivo Oxygen Dynamics Using Plant N-Terminal Degrons in Saccharomyces cerevisiae.- Osmium Resistant Fluorescent Proteins and In-Resin Correlative Light-Electron Microscopy of Epon-Embedded Mammalian Cultured Cells.- Fluorescence-Detection Size-Exclusion Chromatography-Based Thermostability Assay for Membrane Proteins.- Determining Protein-Protein Interaction with GFP-Trap Beads.
