This detailed volume explores protocols for studying the many facets of Ca2+-imaging, Ca2+-signaling, and Ca2+-binding along with background information on the principles and application of these techniques. The content of the book delves into 48 chapters including subjects such as data analysis and modern technologies to study calcium-binding and signaling in cells, the superfamily of calcium-binding proteins characterized by the EF-hand structural motif, as well as their use as diagnostic and prognostic biomarkers in Laboratory Medicine and novel therapeutic drug targets. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Includes cutting-edge techniques

Using Data Mining to Explore Calmodulin Bibliography.- Deep Two-Photon Imaging In Vivo with a Red-Shifted Calcium Indicator.- High Resolution Calcium Imaging Method for Local Calcium Signaling.- Measurement of Contractility and Calcium Release in Cardiac Spheroids.- Simultaneous Recording of Subcellular Ca2+ Signals from the Cytosol and Sarco/Endoplasmatic Reticulum: Compartmentalized Dye Loading, Imaging, and Analysis.- The Use of Complementary Luminescent and Fluorescent Techniques for Imaging Ca2+ Signalling Events during the Early Development of Zebrafish (Danio rerio).- Cellular Ca2+-Responding Nanoluciferase Reporter Gene System Directed by Tandemly Repeated Pseudo-Palindromic NFAT-Response Elements.- Designing Calcium Binding Proteins for Molecular MR Imaging.- Coordination to Divalent Cations by Calcium Binding Proteins.- Chaperoning against Amyloid Aggregation: Monitoring In Vitro and In Vivo.- Ca2+-Binding Proteins of theEF-Hand Superfamily: Diagnostic and Prognostic Biomarkers and Novel Therapeutic Targets.- Gene Transfer of Calcium Binding Proteins into Adult Cardiac Myocytes.- Expression and Purification of Calmodulin for NMR and Other Biophysical Applications.- The Use of Cre-loxP Inducible Mouse Models to Dissect the Specific Roles of Calcineurin Signalling in Myeloid Cells.- Calpain Purification through Calpastatin and Calcium: Strategy and Procedures.- Characterization of the EF-Hand Calcium-Binding Domains of Human Plastins.- Expression and Characterization of MICU2, a Ca2+ Sensor Protein.- S100 Proteins in the Innate Immune Response to Pathogens.- Targeting S100 Calcium-Binding Proteins with Small Molecule Inhibitors.- Monitoring Interactions between S100B and the Dopamine D2 Receptor Using NMR Spectroscopy.- Isolation and Characterization of S100 Protein-Protein Complexes.- The Multifaceted S100A4 Protein in Cancer and Inflammation.- Interaction of S100A6 with Target Proteins In Vitro and in Living Cells.- Preparation of the Oxidized and Reduced Forms of Psoriasin (S100A7).- Preparation and Iron Redox Speciation Study of the Fe(II)-Binding Antimicrobial Protein Calprotectin.- Structural Analysis of S100A8 Complex with Zinc and Calcium: A General Protocol for the Study of S100 Proteins in the Presence of Divalent Cations by X-Ray Crystallography.- Analysis of Ca2+-Dependent Weibel-Palade Body Tethering by Live Cell TIRF Microscopy: Involvement of a Munc13-4/S100A10/Annexin A2 Complex.- Analysis of S100A11 in DNA Damage Repair.- Fluorine-18 Labeling of S100 Proteins for Small Animal Positron Emission Tomography.- Reviewing the Crystal Structure of S100Z and Other Members of the S100 Family: Implications in Calcium-Regulated Quaternary Structure.- High Sensitive Quantitative Binding Assays Using a Nanoluciferase-Fused Probe for Analysis of ALG-2-Interacting Proteins.- Calcium-Induced Protein Folding in Calumenin and Calmodulin.- Measuring CalumeninImpact on ER-Calcium Depletion Using Transient Calumenin Overexpression and Silencing.- Secretagogin Purification and Quality Control Strategies for Biophysical and Cell Biological Studies.- Tryptophan Scanning Mutagenesis of EF-Hand Motifs.- Mapping Calcium-Sensitive Regions in GCAPs by Site-Specific Fluorescence Labeling.- Quantification of Human Swiprosin-1/EFhd2 Expression on Protein and RNA Level.- Three-Dimensional Reconstruction Imaging Method to Study the Function of EFHD2 in Invadopodia Formation.- Characterization of Calcium-Binding Proteins from Parasitic Worms.- High-Sensitivity Troponin Assays in Clinical Diagnostics of Acute Coronary Syndrome.- Targeted Mass Spectrometry of S100 Proteins.- Clinical Use of the Calcium-Binding S100B Protein, a Biomarker for Head Injury.- Serum S100B Levels in Melanoma.- The Ca2+-Binding S100B Protein: An Important Diagnostic and Prognostic Neurobiomarker in Pediatric Laboratory Medicine.- S100A7 in Psoriasis: Immunodetection andActivation by CRISPR.- S100A8/A9 in Myocardial Infarction.- Enzyme-Linked Immunosorbent Assay to Measure S100A12 in Fecal Samples of Children and Adults.- S100 Proteins as Biomarkers in Risk Estimations for Malignant Transformation in Oral Lesions.