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In Fluorescent Protein-Based Biosensors: Methods and Protocols, experts in the field have assembled a series of protocols describing several methods in which fluorescent protein-based reporters can be used to gain unique insights into the regulation of cellular signal transduction. Genetically encodable fluorescent biosensors have allowed researchers to observe biochemical processes within the endogenous cellular environment with unprecedented spatiotemporal resolution. As the number and diversity of available biosensors grows, it is increasingly important to equip researchers with an understanding of the key concepts underlying the design and application of genetically encodable fluorescent biosensors to live cell imaging. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Fluorescent Protein-Based Biosensors: Methods and Protocols promises to be a valuable resource for researchers interested in applying current biosensors to the study of biochemical processes in living cells as well as those interested in developing novel biosensors to visualize other cellular phenomena.
List of contents
The Design and Application of Genetically-Encodable Biosensors Based on Fluorescent Proteins.- An Introduction to Fluorescence Imaging Techniques Geared Towards Biosensor Applications.- Quantitative Measurement of Ca2+ and Zn2+ in Mammalian Cells using Genetically Encoded Fluorescent Biosensors.- Detecting cAMP with an Epac-Based FRET Sensor in Single Living Cells.- Analysis of Compartmentalized cAMP: A Method to Compare Signals from Differently Targeted FRET Reporters.- Genetically Encoded Fluorescent Biosensors for Live-cell Imaging of Lipid Dynamics.- Live Cell Imaging of Cytosolic NADH-NAD+ Redox State using a Genetically Encoded Fluorescent Biosensor.- Measuring Membrane Voltage with Microbial Rhodopsins.- Imaging the Activity of Ras superfamily GTPase Proteins in Small Subcellular Compartments in Neurons.- Imaging Kinase Activity at Protein Scaffolds.- Using a Genetically Encoded FRET-Based Reporter to Visualize calcineurin phosphatase Activity in Living Cells.- Genetically Encoded FRET Indicators for Live-Cell Imaging of Histone Acetylation.- Genetically Encoded Fluorescent Biosensors for Live-Cell Imaging of MT1-MMP Protease Activity.- Biosensor Imaging in Brain Slice Preparations.- Optical Calcium Imaging Using DNA-Encoded Fluorescence Sensors in Transgenic Fruit Flies, Drosophila melanogaster.- Multi-Parameter Live Cell Imaging Approach to Monitor Cyclic AMP and Protein Kinase A Dynamics in Parallel.- FRET and BRET-Based Biosensors in Live Cell Compound Screens.- Integrating Fluorescent Biosensor Data Using Computational Models,
Summary
In Fluorescent Protein-Based Biosensors: Methods and Protocols, experts in the field have assembled a series of protocols describing several methods in which fluorescent protein-based reporters can be used to gain unique insights into the regulation of cellular signal transduction. Genetically encodable fluorescent biosensors have allowed researchers to observe biochemical processes within the endogenous cellular environment with unprecedented spatiotemporal resolution. As the number and diversity of available biosensors grows, it is increasingly important to equip researchers with an understanding of the key concepts underlying the design and application of genetically encodable fluorescent biosensors to live cell imaging. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Fluorescent Protein-Based Biosensors: Methods and Protocols promises to be a valuable resource for researchers interested in applying current biosensors to the study of biochemical processes in living cells as well as those interested in developing novel biosensors to visualize other cellular phenomena.
