Fluorescent Protein-Based Biosensors: Methods and Protocols

ISBN-13: 9781627036214 / Angielski / Twarda / 2013 / 251 str.

Jin Zhang; Qiang Ni; Robert H. Newman

Opis

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.

Spis treści

1. The Design and Application of Genetically-Encodable Biosensors Based on Fluorescent Proteins

Robert H. Newman and Jin Zhang

 

2. An Introduction to Fluorescence Imaging Techniques Geared Towards Biosensor Applications

J. Goedhart, Mark A. Hink, and Kees Jalink

 

3. Quantitative Measurement of Ca2+ and Zn2+ in Mammalian Cells using Genetically Encoded Fluorescent Biosensors

J. Genevieve Park and Amy E. Palmer

 

4. Detecting cAMP with an Epac-Based FRET Sensor in Single Living Cells

J. Klarenbeek and Kees Jalink

 

5. Analysis of Compartmentalized cAMP: A Method to Compare Signals from Differently Targeted FRET Reporters

Alessandra Stangherlin, Andreas Koschinski, Anna Terrin, Anna Zoccarato, He Jiang, Laura Ashley Fields, and Manuela Zaccolo

 

6. Genetically Encoded Fluorescent Biosensors for Live-cell Imaging of Lipid Dynamics

Moritoshi Sato

 

7. Live Cell Imaging of Cytosolic NADH-NAD+ Redox State using a Genetically Encoded Fluorescent Biosensor

Yin Pun Hung and Gary Yellen

 

8. Measuring Membrane Voltage with Microbial Rhodopsins

Daniel R. Hochbaum and Adam Cohen

 

9. Imaging the Activity of Ras superfamily GTPase Proteins in Small Subcellular Compartments in Neurons

Ana F. Oliveira and Ryohei Yasuda

 

10. Imaging Kinase Activity at Protein Scaffolds

Maya T. Kunkel and Alexandra C. Newton

 

11. Using a Genetically Encoded FRET-Based Reporter to Visualize calcineurin phosphatase Activity in Living Cells

Sohum Meta and Jin Zhang

 

12. Genetically Encoded FRET Indicators for Live-Cell Imaging of Histone Acetylation

Kazuki Sasaki and Minoru Yoshida

 

13. Genetically Encoded Fluorescent Biosensors for Live-Cell Imaging of MT1-MMP Protease Activity

Mingxing Ouyang, Shaoying Lu, and Yingxiao Wang

 

14. Biosensor Imaging in Brain Slice Preparations

Marina Polito, Pierre Vincent, and Elvire Guiot

 

15. Optical Calcium Imaging Using DNA-Encoded Fluorescence Sensors in Transgenic Fruit Flies, Drosophila melanogaster

Shubham Dipt, Thomas Riemensperger, and André Fiala

 

16. Multi-Parameter Live Cell Imaging Approach to Monitor Cyclic AMP and Protein Kinase A Dynamics in Parallel

Nwe-Nwe Aye-Han and Jin Zhang

 

17. FRET and BRET-Based Biosensors in Live Cell Compound Screens

Katie Herbst-Robinson, Jessica Yang, and Jin Zhang

 

18. Integrating Fluorescent Biosensor Data Using Computational Models

Eric C. Greenwald, Renata K. Polanowska-Grabowska, and Jeffery J. Saucerman