Calcium Signaling Protocols

Part I: General

1. Fluorescent Measurement of [Ca^2]]_c: Basic Practical Considerations

Alec W.M. Simpson

2. Measurement of [Ca^2]]_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 [Ca^2]]_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 [Ca^2]]_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 Calì, 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