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This extensively updated and expanded second edition contains new chapters on the elctrophysiological 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 dacets of Ca2+ signaling. The protocols follow the successful Methods in Molecular BiologyT series format, each offering 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.
Part I. General; Fluorescent Measurement of [Ca2+]c: Basic Practical Considerations; Alec W. M. Simpson; Measurement of [Ca2+]i in Whole Cell Suspensions Using Fura-2; Robert A. Hirst, Charlotte Harrison, Kazuyoshi Hirota, and David G. Lambert; Measurement of [Ca2+]i in Cell Suspensions Using Indo-1; Adriaan Nelemans; Part II. Specialist Measurement Systems; Confocal Microscopy: Theory and Applications for Cellular Signaling; Stephen C. Tovey, Paul J. Brighton, and Gary B. Willars; Single Cell and Subcellular Measurement of Intracellular Ca2+ Concentration ([Ca2+]i); Anthony J. Morgan and Andrew P. Thomas; Ratiometric Ca2+ Measurements Using the FlexStation[registered] Scanning Fluorometer; Ian C. B. Marshall, Izzy Boyfield, and Shaun McNulty; Measuring Ca2+ Changes in Multiwell Format Using the Fluorometric Imaging Plate Reader (FLIPR[registered]); Ian C. B. Marshall, Davina E. Owen, and Shaun McNulty; Part III. Measurement of Ca 2+ Channel Activity; Measurement of Ca2+ Entry into Cells Using 45Ca 2+; Mercedes Villarroya, Manuela G. Lopez, Maria F. Cano-Abad, and Antonio G. Garcia; Measurement of [3H]PN200-110 and w-Conotoxin MVIIA Binding; Kazuyoshi Hirota and David G. Lambert; Whole-Cell Patch Clamp Recording of Voltage-Sensitive Ca 2+ Channel Currents in Single Cells: Heterologous Expression Systems and Dissociated Brain Neurons; Atticus H. Hainsworth, Andrew D. Randall, and Alessandro Stefani; Part IV. Measurement of Ins(1,4,5)P3 and Ca2+ Release from Intracellular Stores; Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol Labeling Protocols in Permeabilized Cells; Alison Skippen, Philip Swigart, and Shamshad Cockcroft.; Measurement of Inositol(1,4,5)triphosphate Using a Stereospecific Radioreceptor Mass Assay; Darren Smart; Measurement of Calcium Fluxes in Permeabilised Cells Using a 45Ca 2+ Uptake and Release Assay; Robert A. Wilcox; Microinjection of myo-inositol(1,4,5)trisphosphate and Other Calcium-Mobilizing Agents into Intact Adherent Cells; Robert A. Wilcox, Ian D. Forsythe, and Terence J. McCann; Measurement of Free [Ca2+] Changes in Agonist-Sensitive Internal Stores Using Compartmentalized Fluorescent Indicators; Aldebaran M. Hofer. Part V. Specialist Measurement Techniques; Measurement of [Ca2+]i in Smooth Muscle Strips Using Front-Surface Fluorimetry; Hideo Kanaide; Measurement of Calcium and Movement in Heart Cells; Leong L. Ng and Paulene A. Quinn; Simultaneous Analysis of Intracellular pH and Ca2+ from Cell Populations; Raul Martinez-Zaguilan, Linda S. Tompkins, Robert J. Gillies, and Ronald M. Lynch; Measurement of Cytosolic-Free Ca2+ in Plant Tissue; Martin R. McAinsh and Carl K.-Y Ng; Part VI. Ca2+ Sensitive Targets; Assay and Purification of Calmodulin-Dependent Protein Kinase; Rajendra K. Sharma; Assay and Purification of Calmodulin-Dependent Cyclic Nucleotide Phosphodiesterase and Isozyme Separation; Rajendra K. Sharma; Measurement of Ca2+-ATPase activity (in PMCA and SERCA1); Danuta Kosk-Kosicka.
: "...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