Calcium Signaling Protocols

1. Front Matter

   Pages i-x

   PDF

2. General

   1. Front Matter

      Pages 1-1

      PDF

   2. Fluorescent Measurement of [Ca2+]c

      • Alec W. M. Simpson

      Pages 3-36

   3. Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2

      • Robert A. Hirst, Charlotte Harrison, Kazuyoshi Hirota, David G. Lambert

      Pages 37-45

   4. Measurement of [Ca2+]i in Cell Suspensions Using Indo-1

      • Adriaan Nelemans

      Pages 47-53

3. Specialist Measurement Systems

   1. Front Matter

      Pages 55-55

      PDF

   2. Confocal Microscopy

      • Stephen C. Tovey, Paul J. Brighton, Gary B. Willars

      Pages 57-85

   3. Single-Cell and Subcellular Measurement of Intracellular Ca2+ Concentration

      • Anthony J. Morgan, Andrew P. Thomas

      Pages 87-117

   4. Ratiometric Ca2+ Measurements Using the FlexStation® Scanning Fluorometer

      • Ian C. B. Marshall, Izzy Boyfield, Shaun McNulty

      Pages 119-124

   5. Measuring Ca2+ Changes in Multiwell Format Using the Fluorometric Imaging Plate Reader

      • Ian C. B. Marshall, Davina E. Owen, Shaun McNulty

      Pages 125-131

4. Measurement of Ca2+ Channel Activity

   1. Front Matter

      Pages 133-133

      PDF

   2. Measurement of Ca2+ Entry Using 45Ca2+

      • Mercedes Villarroya, Manuela G. López, María F. Cano-Abad, Antonio G. García

      Pages 135-145

   3. Measurement of [3H]PN200-110 and [125I]ω-Conotoxin MVIIA Binding

      • Kazuyoshi Hirota, David G. Lambert

      Pages 147-159

   4. Whole-Cell Patch Clamp Recording of Voltage-Sensitive Ca2+ Channel Currents

      • Atticus H. Hainsworth, Andrew D. Randall, Alessandro Stefani

      Pages 161-179

5. Measurement of Ins(1,4,5,)P3 and Ca2+ Release From Intracellular Stores

   1. Front Matter

      Pages 181-181

      PDF

   2. Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol-Labeling Protocols in Permeabilized Cells

      • Alison Skippen, Philip Swigart, Shamshad Cockcroft

      Pages 183-193

   3. Measurement of Inositol(1,4,5)Trisphosphate Using a Stereospecific Radioreceptor Mass Assay

      • Darren Smart

      Pages 195-203

   4. Measurement of Calcium Fluxes in Permeabilized Cells Using a 45Ca2+ Uptake and Release Assay

      • Robert A. Wilcox

      Pages 205-212

   5. Microinjection of myo-Inositol(1,4,5)Trisphosphate and Other Calcium-Mobilizing Agents Into Intact Adherent Cells

      • Robert A. Wilcox, Ian D. Forsythe, Terence J. McCann

      Pages 213-228

   6. Measurement of Free [Ca2+] Changes in Agonist-Sensitive Internal Stores Using Compartmentalized Fluorescent Indicators

      • Aldebaran M. Hofer

      Pages 229-247

In the first edition of Calcium Signaling Protocols I began by writing “The regula- 2+ tion of intracellular Ca is a common theme presented in many papers over the last 20 2+ or so years and the description of the Ca -sensitive indicator dye fura-2 in 1985 resulted in a massive increase in these types of studies. ” This statement is as true in 2005 as it was in 1999, but 20 or so years is now 30 years! There has been some reorganization of the volume such that there are now 22 ch- ters including five new ones, all written by experts in their field. These new chapters 2+ include use of the FlexStation and electrophysiological measurement of Ca channel activity. The book is broken into six parts. Part I is a general coverage of basic theory and the simplest use of fluorescent indicators. Part II covers specialist measurement 2+ systems and Part III covers measurement of Ca channel activity. Assessment of 2+ release of stored Ca is covered in some detail in Part IV, with Parts V and VI cover- 2+ ing specialist measurement techniques and Ca -sensitive targets. Putting a book like this together, even as a second edition, takes time and I am, again, indebted to the individual authors for their help and patience. I am also very grateful to Professor John M. Walker, the series editor, for his continued help and advice over the course of this project.

• ATP
• ATPase
• Calcium
• Endoplasmatisches Reticulum
• Nucleotide
• molecular biology
• protein
• reproducible techniques

From Reviews of the First Edition...

"...will find a place on the bench in any lab that is moving into calcium signaling, or expanding its repertoire of techniques in calcium signaling. It will be valuable as a resource for...years to come." -Cell Biology International

"This book achieves much...For anyone remotely interested in Ca2+ it will be a very valuable resource in their lab" -Quarterly Review of Biology

". . .recommended to investigators interested in calcium measurement using optical methods. "-Doody's Health Sciences Book Review Journal

"Most protocols identify required materials such as buffers, reagents and cell lines. Methods are presented in a step-by-step format. Hopefully these protocols are adaptable by scientists for use in specific research. Certainly, the methods are specific enough not to leave the researcher stranded. Calcium Signaling Protocols is recommended for academic health and science libraries who support academic research."-E-Streams

• Department of Cardiovascular Sciences, Division of Anaesthesia, Critical Care, and Pain Management, Leicester Royal Infirmary, Leicester, UK

  David G. Lambert

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