G protein-coupled receptors. Part A ([2017])

• Record Type: Book
• Further Information: Note: Edited by Arun K. Shukla.
• Editors: Shukla, Arun K
• Contents: Front Cover; G Protein-Coupled Receptors Part A; Copyright; Contents; Contributors; Preface; Chapter 1: Measurement of GPCR-G protein activity in living cells; 1. Introduction; 2. Materials; 2.1. Generations of Sensors; 2.2. Cell Culture; 2.3. Transfection of Cells; 2.4. Stable Cell Line Generation; 2.5. Live-Cell Imaging; 2.6. Fluorescence Imaging Plate Reader Assay; 3. Methods; 3.1. Generations of Sensors Using Gibson Assembly Protocol; 3.2. Cell Culture; 3.2.1. HeLa Cell-Specific Growth Media Preparation; 3.2.2. Passaging cells 2.1.1. ALPHAscreen (Amplified Luminescent Proximity Homogenous Assay)2.1.2. DELFIA (Dissociation-Enhanced Lanthanide Fluorescence Immunoassay); 2.1.3. HTRF (Homogenous Time Resolved Fluorescence); 2.1.4. Fluorescence polarization assay; 2.2. Luminescence-Based cAMP Measuring Assays; 2.2.1. CRE-luciferase reporter gene assay; 2.2.2. GloSensor assay; 3. Reagents and Consumables; 4. Methods; 4.1. Cell Culture Maintenance and Trypsinization; 4.2. Transient Transfection of HEK293T Cells; 4.3. Preparation of Transfected Cell Plate; 4.4. Test Compound Dilution for GloSensor Assay 4.5. Test Compound Addition to the Cell Plate4.6. Compound Addition to Screen Ligands for GS-Coupled Receptors; 4.7. Compound Addition to Screen Ligands for Gi-Coupled Receptors; 5. Data Analysis; 5.1. Data Analysis for Agonist Activity; 5.2. Data Analysis for Antagonist Activity; 6. Results and Data Interpretation; 7. Final Comments; Acknowledgments; References; Chapter 3: cAMP assays in GPCR drug discovery; 1. Introduction; 2. Materials; 2.1. Cell Culture; 3. Method; 3.1. Troubleshooting; 3.2. Notes; References Chapter 4: Measurement of intracellular Ca2+ mobilization to study GPCR signal transduction1. Introduction; 2. Materials; 2.1. Maintenance of Cell Lines; 2.2. Generation of "Stable Cell Lines"; 2.3. Fura2AM Loading and Spectrophotometric Analysis; 3. Methods; 3.1. Maintenance of Cell Lines; 3.2. Generation of "Stable Cell Lines"; 3.3. Fura2AM Loading; 3.4. Spectrophotometric Analysis; 4. Notes; References; Chapter 5: Visualization and quantification of GPCR trafficking in mammalian cells by confocal microscopy; 1. Introduction; 2. Methods; 2.1. Required Materials
• Publisher Details: Cambridge, MA, USA : Academic Press is an imprint of Elsevier
• Publication Date: 2017
• Copyright Date: 2017
• Extent: 1 online resource, color illustrations
• Subjects: 571.6; Cytology -- Technique; Cell receptors; SCIENCE / Life Sciences / Anatomy & Physiology; Cell receptors; Cytology -- Technique; GTP-Binding Proteins; Cytological Techniques; Receptors, Cell Surface; Electronic books
• Languages: English
• ISBNs: 9780128133200; 0128133201
• Related ISBNs: 9780128133194; 9780128133194
• Notes: Note: Includes bibliographical references and index.; Note: Print version record.
• Access Rights: Legal Deposit; Only available on premises controlled by the deposit library and to one user at any one time; The Legal Deposit Libraries (Non-Print Works) Regulations (UK).
• Access Usage: Restricted: Printing from this resource is governed by The Legal Deposit Libraries (Non-Print Works) Regulations (UK) and UK copyright law currently in force.