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Fluorescent proteins
Title:
Fluorescent proteins
Author:
Sullivan, Kevin Francis, 1955-
ISBN:
9780080557243
9780123725585
9781281144768
9786611144760
Edition:
2nd ed.
Publication Information:
Amsterdam ; London : Elsevier/Academic Press, ©2008.
Physical Description:
1 online resource (xx, 592 pages, 48 unnumbered pages of plates) : illustrations
Series:
Methods in cell biology, v. 85
Methods in cell biology ; v. 85.
Contents:
Front Cover; Methods in Cell Biology; Copyright Page; Contents; Contributors; Preface; Chapter 1: Autofluorescent Proteins; A. Structure; B. Stability, Folding, and Multimerization; C. Spectra and Photophysical Dynamics; A. Multiple Labeling; B. Dynamic Imaging; C. Protein-Protein Interactions; References; Chapter 2: Functional Fusion Proteins by Random Transposon-Based GFP Insertion; B. The Transposition Reaction; C. Transformation Requirements and Troubleshooting; D.E.coli Colony Selection and Growth in a 96-Well Format; E. Backing Up the Experiment: Making 10% Glycerol Stocks.
F. 96-Well Mini-Preparation Purification of Plasmid DNAG. Preparation of HEK 293 Cells; H. Transient Transfection of HEK 293 Cells in a 96-Well Format; I. Screening Live HEK 293 Cells for GEP Fluorescence; J. Removing the Selection Cassette with Restriction Digestion and Re-Ligation; IV. Materials; A. PCR Amplification of the Transposon; B. The Transposition Reaction; C. Transformation Requirements and Troubleshooting; D.E.coli Colony Selection and Growth in a 96-Well Format; E. Backing Up the Experiment: Making 10% Glycerol Stocks; F. 96-Well-Mini-Preparation Purification of Plasmid DNA.
G. Preparation of HEK 293 CellsH. Transient Transfection of HEK 293 Cells in a 96-Well Format; I. Screening Live HEK 293 Cells for GEP Fluorescence; J. Removing the Selection Cassette with Restriction Digestion and Re-Ligation; V. Discussion; Acknowledgments; References; Chapter 3: Fluorescent Proteins for Photoactivation Experiments; A. Photoactivatable Fluorescent Proteins: Aequorea victoria GFP; B. Photoactivatable Fluorescent Proteins: DsRed Fluorescent Protein; C. Photoactivatable Fluorescent Proteins: Green-to-Red Photoconversions.
D. Photoactivatable Fluorescent Proteins: Cyan-to-Green PhotoconversionE. Photoactivatable Fluorescent Proteins: Reversible; A. Protein Dynamics; B. Fluorescence Pulse-Labeling.; C. Photoquenching Fluorescence Resonance Energy Transfer; D. Photoactivated Localization Microscopy; References; Chapter 4: Design and Optimization of Genetically Encoded Fluorescent Biosensors: GTPase Biosensors; B. Expression of the Biosensor in Cells; A. DNA Sequence for the pTriEX-4-Biosensor Construct; VIII. Appendix II; A. Media Formulation for Ham's F-12K Phenol Red-Free; References.
Chapter 5: Fast 4D MicroscopyA. Imaging Modes and Combination with Other Functionalities into a Multifunctional System; B. Keeping the Sampled Volume Immobile and Test for It; C. The Impact of Optical Blur, Noise, Aberrations, and Calibration Defects; D. Setting Up a Rapid 4D Acquisition; IV. Conclusions; Acknowledgments; References; Chapter 6: Single-Molecule Imaging of Fluorescent Proteins; References; Chapter 7: Counting Kinetochore Protein Numbers in Budding Yeast Using Genetically Encoded Fluorescent Proteins; III. Sample Preparation; IV. Microscope and Image Acquisition System.
Abstract:
This new edition of Fluorescent Proteins presents current applications of autofluorescent proteins in cell and molecular biology authored by researchers from many of the key laboratories in the field. Starting from a current review of the broad palette of fluorescent proteins available, several chapters focus on key autofluorescent protein variants, including spectral variants, photodynamic variants as well as chimeric FP approaches. Molecular applications are addressed in chapters that detail work with single molecules, approaches to generating protein fusions and biosensors as well as analysis of protein-protein interactions in vivo by FRET, fluorescence polarization and fluorescence cross corrlation techniques. A number of approaches to in vivo dynamics are presented, including FRAP, photoactivation, and 4-dimensional microscopy. Behavior of spindle components, membrane proteins, mRNA trafficking as well as analysis of cell types in tissues and in development are detailed and provide models for a wide variety of experimental approaches. In addition, several chapters deal directly with the computational issues involved in processing multidimensional image data and using fluorescent imaging to probe cellular behavior with quantitative modeling. This volume brings together the latest perspective and techniques on fluorescent proteins and will be an invaluable reference in a wide range of laboratories.
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Electronic Access:
ScienceDirect https://www.sciencedirect.com/science/book/9780123725585 ScienceDirect https://www.sciencedirect.com/science/bookseries/0091679X/85Available:*
Shelf Number | Item Barcode | Shelf Location | Status |
|---|---|---|---|
| QH581.2 .F56 2008 EB | 1189283-1001 | Elsevier E-Book Collections | Searching... |
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