| Z704717 Sigma |
Calcium Signaling Protocols, 2nd ed. |
General description
This extensively updated and expanded second edition contains new chapters on the electrophysiological measurement of Ca2+ channel activity, enhanced coverage of confocal microscopy, and practical tutorials on two of the most common industrial high-throughput machines, the Fluormetric Imaging Plate Reader (FLIPR), and the FlexStation. All experimental chapters provide readily reproducible techniques for measuring the many facets of Ca2+ signaling. The protocols offer step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Table of Contents
PART I. GENERAL
1. Fluorescent Measurement of [Ca2+]c: Basic Practical Considerations
2. Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2
3. Measurement of [Ca2+]i in Cell Suspensions Using Indo-1
PART II. SPECIALIST MEASUREMENT SYSTEMS
4. Confocal Microscopy: theory and Applications for Cellular Signaling
5. Single Cell and Subcellular Measurement of Intracellular Ca2+ Concentration
6. Ratiometric Ca2+ Measurements Using the FlexStation Scanning Fluorimeter
7. Measuring Ca2+ Changes in Multiwell Format Using the Fluorimetric Imaging Plate Reader
PART III. MEASUREMENT OF Ca2+ CHANNEL ACTIVITY
8. Measurement of Ca2+ Entry Using 45Ca2+
9. Measurement of [3H]PN200-110 and [125]ω-Conotoxin MVIIA Binding
10. Whole Cell Patch Clamp Recording of Voltage Sensitive Ca2+ Channel Currents: Heterologous Expression Systems and Dissociated Brain Neurons
PART IV. MEASUREMENT OF Ins(1,4,5)P3 and Ca2+ RELEASE FROM INTRACELLULAR STORES
11. Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H] Inositol Labeling Protocols in Permeabilized Cells
12. Measurement of Inositol(1,4,5)Triphosphate Using a Stereospecific Radioreceptor Mass Assay
13. Measurement of Calcium Fluxes in Permeabilized Cells Using a 45Ca2+ Uptake and Release Assay
14. Micorinjection of myo-Inositol(1,4,5)Triphosphate and Other Calcium Mobilizing Agents Into Intact Adherent Cells
15. Measurement of Free[Ca2+] Changes in Agonist Sensitive Internal Stores Using Compartmentalized Fluorescent Indicators
PART V. SPECIALIST MEASUREMENT TECHNIQUES
16. Measurement of [Ca2+]i in Smooth Muscle Strips Using Front Surface Fluorimetry
17. Measurement of Calcium and Movement in Heart Cells
18. Simultaneous Analysis of Intracellular pH and Ca2+ From Cell Populations
19. Measurement of Cytosolic Free Ca2+ in Plant Tissue
PART VI. Ca2+ SENSITIVE TARGETS
20. Assay and Purification of Calmodulin Dependent Protein Kinase
21. Assay and Purification of Calmodulin Dependent Cyclic Nucleotide Phosphsdiesterase and Isozyme Separation
22. Measurement of Ca2+ ATPase Activity (in PMCAand SERCA1)
Index
1. Fluorescent Measurement of [Ca2+]c: Basic Practical Considerations
2. Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2
3. Measurement of [Ca2+]i in Cell Suspensions Using Indo-1
PART II. SPECIALIST MEASUREMENT SYSTEMS
4. Confocal Microscopy: theory and Applications for Cellular Signaling
5. Single Cell and Subcellular Measurement of Intracellular Ca2+ Concentration
6. Ratiometric Ca2+ Measurements Using the FlexStation Scanning Fluorimeter
7. Measuring Ca2+ Changes in Multiwell Format Using the Fluorimetric Imaging Plate Reader
PART III. MEASUREMENT OF Ca2+ CHANNEL ACTIVITY
8. Measurement of Ca2+ Entry Using 45Ca2+
9. Measurement of [3H]PN200-110 and [125]ω-Conotoxin MVIIA Binding
10. Whole Cell Patch Clamp Recording of Voltage Sensitive Ca2+ Channel Currents: Heterologous Expression Systems and Dissociated Brain Neurons
PART IV. MEASUREMENT OF Ins(1,4,5)P3 and Ca2+ RELEASE FROM INTRACELLULAR STORES
11. Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H] Inositol Labeling Protocols in Permeabilized Cells
12. Measurement of Inositol(1,4,5)Triphosphate Using a Stereospecific Radioreceptor Mass Assay
13. Measurement of Calcium Fluxes in Permeabilized Cells Using a 45Ca2+ Uptake and Release Assay
14. Micorinjection of myo-Inositol(1,4,5)Triphosphate and Other Calcium Mobilizing Agents Into Intact Adherent Cells
15. Measurement of Free[Ca2+] Changes in Agonist Sensitive Internal Stores Using Compartmentalized Fluorescent Indicators
PART V. SPECIALIST MEASUREMENT TECHNIQUES
16. Measurement of [Ca2+]i in Smooth Muscle Strips Using Front Surface Fluorimetry
17. Measurement of Calcium and Movement in Heart Cells
18. Simultaneous Analysis of Intracellular pH and Ca2+ From Cell Populations
19. Measurement of Cytosolic Free Ca2+ in Plant Tissue
PART VI. Ca2+ SENSITIVE TARGETS
20. Assay and Purification of Calmodulin Dependent Protein Kinase
21. Assay and Purification of Calmodulin Dependent Cyclic Nucleotide Phosphsdiesterase and Isozyme Separation
22. Measurement of Ca2+ ATPase Activity (in PMCAand SERCA1)
Index
Related Information
Links
Page Options| |
|
© 2008 Sigma-Aldrich Co. Reproduction forbidden without permission. |