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Understanding cellular function is a pivotal aim in the life sciences. Monitoring the interplay of regulatory proteins illuminates cellular function on a molecular level. One powerful method is fluorescence microscopy, which enables the spatio-temporal resolution of cellular events. To this end, several fluorescent reporters can be monitored within a single cell and interdependency between these factors can be determined. This book presents fluorescent reporters for monitoring protein kinase C, a central actor on the stage of cellular signalling. Aspects ranging from probe design to the evaluation of experimental results are discussed. Further, experiments designed for high-throughput screening of new probe variants or PKC-effector molecules are described, including methods for visualizing and judging the observed data. Finally, the probe''s capacity for use in multiparameter experiments is demonstrated. This book is complemented by a detailed experimental part including a flexible cloning system for rapid probe development and computer scripts supporting the analysis of experimental data. This book is directed to researchers interested in fluorescence probe development.
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