The regulation of intracellular Ca2+ has continued to be a powerful area of study since the publication of the first and second editions of Calcium Signaling Protocols, and the developments in the field have also, naturally, continued. With the third edition, expert contributors explore some of the exciting new molecular techniques that have both enabled new studies of intracellular Ca2+ regulation and provided much new information on processes.
Comprised of five main section, Calcium Signaling Protocols covers theoretical and very simple suspension-based fluorimetric assays, specialist measurement systems, measurement of channel activity, measurement of store release, as well as specialist measurement techniques which include targeted probes, using G-protein chimeras to force Ca2+ signalling for screening, and genetically encoded sensors.
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 tips on troubleshooting and avoiding known pitfalls. Updated and accessible, Calcium Signaling Protocols: Third Edition will be a valuable reference for all those contemplating a move into the study of intracellular Ca2+.
Part I: General
1. Fluorescent Measurement of [Ca2+]c: Basic Practical Considerations Alec W.M. Simpson
2. Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2 Anish Patel, Robert A. Hirst, Charlotte Harrison, Kazuyoshi Hirota, and David G. Lambert
Part II: Specialist Measurement Systems
3. Confocal Microscopy: Theory and Applications for Cellular Signaling Stephen C. Tovey, Paul J. Brighton, Edward T.W. Bampton, Yan Huang, and Gary B. Willars
4. Ratiometric Ca2+ Measurements Using the FlexStation(R) Scanning Fluorometer Ian C.B. Marshall, Izzy Boyfield, and Shaun McNulty
5. Measuring Ca2+ Changes in Multiwell Format Using the Fluorometric Imaging Plate Reader (FLIPR(R)) Ian C.B. Marshall, Davina E. Owen, and Shaun McNulty
6. Ratiometric [Ca2+]i Measurements in Adherent Cell-lines Using the NOVOstar Microplate Reader Benjamin D. Hunt and David G. Lambert
Part III: Measurement of Ca2+ Channel Activity
7. Whole-Cell Patch Clamp Recording of Voltage-Sensitive Ca2+ Channel Currents in Single Cells: Heterologous Expression Systems and Neurones Jon Brown, Atticus H. Hainsworth, Alessandro Stefani, and Andrew D. Randall
8. Combined Calcium Fluorescence Recording with Ionic Currents in Contractile Cells Richard D. Rainbow
Part IV: Measurement of Ins(1,4,5)P3 and Ca2+ Release Form Intracellular Stores
9. Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol Labeling Protocols in Permeabilized Cells Alison Skippen, Philip Swigart, and Shamshad Cockcroft
10. Single Cell Imaging Techniques for the Real-Time Detection of IP3 in Live Cells Carl P. Nelson
11. Measurement of Inositol(1,4,5)Trisphosphate Using a Stereospecific Radioreceptor Mass Assay Darren Smart
Part V: Specialist Measurement Techniques
12. Measurement of [Ca2+]i in Smooth Muscle Strips Using Front-Surface Fluorimetry Hideo Kanaide and Katsuya Hirano
13. Calcium Measurements from Whole Heart Using Rhod-2 Bum-Rak Choi
14. Measurement of Changes in Endothelial and Smooth Muscle Ca2+ in Pressurized Arteries Kim A. Dora and Michael A. Hill
15. Single Cell and Subcellular Measurements of Intracellular Ca2+ Concentration John G. McCarron, Marnie L. Olson, Susan Chalmers, and John M. Girkin
16. Simultaneous Analysis of Intracellular pH and Ca2+ from Cell Populations Raul Martinez-Zaguilan, Linda S. Tompkins, Robert J. Gillies, and Ronald M. Lynch
17. Measurements of Ca2+ Concentration with Recombinant Targeted Luminescent Probes Denis Ottolini,Tito Cali, and Marisa Brini
18. Chimeric G-Proteins in Fluorimetric Calcium Assays: Experience with Opioid Receptors Valeria Camarda and Girolamo Calo
19. Compartmentalizing Genetically-Encoded Calcium Sensors David A. Williams, Mastura Monif, and Kate L. Richardson
20. Measurement of Cytosolic-Free Ca2+ in Plant Tissue Martin R. McAinsh and Carl K.-Y. Ng
21. Measurement of Ca2+-ATPase Activity (in PMCA and SERCA1) Danuta Kosk-Kosicka
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