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This detailed volume explores techniques for performing calcium signaling in diverse cell types, alongside detailed methods for data analysis. Beginning with chapters on cellular assays to assess calcium signaling, the book continues with sections on imaging methods, methods to assess calcium within primary cilia, the endoplasmic reticulum, and the mitochondria, as well as tissue-specific calcium signaling methods, including the establishment of primary cell or tissue models and measuring calcium in whole organisms such as Drosophila. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Calcium Signaling: Methods and Protocols serves as an ideal guide for laboratories of all types seeking to measure calcium robustly.
List of contents
Measuring GPCR-Induced Intracellular Calcium Signaling Using a Quantitative High-Throughput Assay.- Quantifying Gq Signaling Using the IP1 Homogenous Time Resolved Fluorescence (HTRF) Assay.- Measuring IP3 Generation in Real-Time Using a NanoBiT Luminescence Biosensor.- Monitoring Calcium-Sensing Receptor (CaSR) Induced Intracellular Calcium Flux Using an Indo-1 Flow Cytometry Assay.- Bioluminescence Resonance Energy Transfer (BRET) Assay to Measure Gq Recruitment to the Ghrelin Receptor.- Analysis of Calcium-Sensing Receptor Signaling Using Dual Luciferase Assays.- Calcium Imaging in Brain Tissue Slices.- Two-Photon Microscopy to Measure Calcium Signaling in the Living Brain.- Measuring Calcium-Sensing Receptor Agonist-Driven Insertional Signaling (ADIS) and Trafficking by TIRF Microscopy.- Measuring Calcium Signaling at the Primary Cilia.- Monitoring ER Ca2+ by Luminescence with Low Affinity GFP-Aequorin Protein (GAP).- Analyzing Mitochondrial Calcium Influx in Isolated Mitochondria.- Measuring Calcium Levels in Bone Resorbing Osteoclasts and Bone Forming Osteoblasts.- Quantitative Intravital Calcium Imaging in Mouse Kidney.- Methods for Imaging Intracellular Calcium Signals in the Mouse Mammary Epithelium in 2- and 3-Dimensions.- Live Cell Calcium Imaging in 3D Intestinal Organoids.- Calcium Imaging and Analysis in Beta Cells in Acute Mouse Pancreas Tissue Slices.- Real-Time Imaging of Calcium Dynamics in Human Sperm after Precise Single-Cell Stimulation.- Calcium Imaging in Drosophila.- Computational Analysis of Calcium Flux Data Using R.
