JoVE Journal
Biology
Biology
A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
Summary
A method for estimating the affinity constant of an agonist for the active state (Kb) of a G protein-coupled receptor is described. The analysis provides absolute or relative measures of Kb depending on whether constitutive receptor activation is measurable. Our method applies to various responses downstream from receptor activation.
Tags
Quantifying Agonist Activity G Protein-coupled Receptors Signaling Pathway Cell Response Tissue Response Single Receptor Analysis Downstream Response Agonist Affinity Constant Active State Inactive State Ligands Receptor Activation Concentration Of Agonist Observed Affinity Constant Efficacy Methods For Analysis
PLAYLIST
The Theory of Receptors and Ligands
-
Education • Core: BiologyG-protein Coupled Receptors -
Education • Core: BiologyInternal Receptors -
Education • Core: BiologyWhat are Second Messengers? -
Education • Core: BiologyIntracellular Signaling Cascades -
Education • Core: BiologyInduced-fit Model -
Education • Core: BiologyEnzyme Inhibition -
Education • Core: BiologyAllosteric Regulation -
Education • Core: BiologyCofactors and Coenzymes -
Research • Immunology and InfectionCo-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
Basic Research Tools
-
Education • Biomedical EngineeringImaging Biological Samples with Optical and Confocal Microscopy -
Education • Basic Methods in Cellular and Molecular BiologyAn Introduction to Transfection -
Education • Cell BiologyThe Transwell Migration Assay -
Research • Bioengineering2D and 3D Matrices to Study Linear Invadosome Formation and Activity -
Research • BiologyAnalysis of Cell Cycle Position in Mammalian Cells -
Education • Cell BiologyCell-surface Biotinylation Assay -
Education • Cell BiologyThe ATP Bioluminescence Assay -
Education • Cell BiologyDetecting Reactive Oxygen Species -
Education • Cell BiologyThe TUNEL Assay -
Research • Cancer ResearchAssessing Cell Viability and Death in 3D Spheroid Cultures of Cancer Cells -
Research • BiologyViability Assays for Cells in Culture
Cell-Based Assays
-
Research • NeuroscienceMethods for the Discovery of Novel Compounds Modulating a Gamma-Aminobutyric Acid Receptor Type A Neurotransmission -
Research • Cancer ResearchDetecting the Ligand-binding Domain Dimerization Activity of Estrogen Receptor Alpha Using the Mammalian Two-Hybrid Assay -
Research • MedicineA Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators -
Research • BiologyA BW Reporter System for Studying Receptor-Ligand Interactions -
Research • MedicineAn Organotypic High Throughput System for Characterization of Drug Sensitivity of Primary Multiple Myeloma Cells -
Research • BiochemistryAn Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects -
Research • BioengineeringTherapeutic Gene Delivery and Transfection in Human Pancreatic Cancer Cells using Epidermal Growth Factor Receptor-targeted Gelatin Nanoparticles
Tissue-Based Testing
-
Research • MedicineBladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology -
Research • BiologyMeasurement of Smooth Muscle Function in the Isolated Tissue Bath-applications to Pharmacology Research -
Research • Immunology and InfectionThe Ex Vivo Culture and Pattern Recognition Receptor Stimulation of Mouse Intestinal Organoids -
Research • MedicineForskolin-induced Swelling in Intestinal Organoids: An In Vitro Assay for Assessing Drug Response in Cystic Fibrosis Patients -
Research • NeuroscienceProbing Nicotinic Acetylcholine Receptor Function in Mouse Brain Slices via Laser Flash Photolysis of Photoactivatable Nicotine -
Research • BioengineeringAutomated Contraction Analysis of Human Engineered Heart Tissue for Cardiac Drug Safety Screening -
Research • NeuroscienceA Human Blood-Brain Interface Model to Study Barrier Crossings by Pathogens or Medicines and Their Interactions with the Brain -
Research • ChemistryAn Intestine/Liver Microphysiological System for Drug Pharmacokinetic and Toxicological Assessment
In Vivo Testing
-
Research • MedicineLive Imaging of Drug Responses in the Tumor Microenvironment in Mouse Models of Breast Cancer -
Research • Immunology and InfectionA Robust Pneumonia Model in Immunocompetent Rodents to Evaluate Antibacterial Efficacy against S. pneumoniae, H. influenzae, K. pneumoniae, P. aeruginosa or A. baumannii -
Research • NeuroscienceSystemic and Local Drug Delivery for Treating Diseases of the Central Nervous System in Rodent Models -
Research • BiologyA Comparative Study of Drug Delivery Methods Targeted to the Mouse Inner Ear: Bullostomy Versus Transtympanic Injection -
Research • Immunology and InfectionThe Mesenteric Lymph Duct Cannulated Rat Model: Application to the Assessment of Intestinal Lymphatic Drug Transport -
Research • MedicineUse of Rabbit Eyes in Pharmacokinetic Studies of Intraocular Drugs -
Research • MedicinePercutaneous Contrast Echocardiography-guided Intramyocardial Injection and Cell Delivery in a Large Preclinical Model -
Research • MedicineA Rabbit Model of Aqueous-Deficient Dry Eye Disease Induced by Concanavalin A Injection into the Lacrimal Glands: Application to Drug Efficacy Studies
Dose-Response Curves
Drug Screening
-
Research • BiochemistryKinase Inhibitor Screening In Self-assembled Human Protein Microarrays -
Research • BiologyAvidity-based Extracellular Interaction Screening (AVEXIS) for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions -
Research • BiologyHigh-throughput Screening for Small-molecule Modulators of Inward Rectifier Potassium Channels -
Research • BiochemistryParallel Interrogation of β-Arrestin2 Recruitment for Ligand Screening on a GPCR-Wide Scale using PRESTO-Tango Assay -
Research • Immunology and InfectionIdentification of Mediators of T-cell Receptor Signaling via the Screening of Chemical Inhibitor Libraries -
Research • Cancer ResearchPhysiologic Patient Derived 3D Spheroids for Anti-neoplastic Drug Screening to Target Cancer Stem Cells -
Research • Bioengineering3D Microtissues for Injectable Regenerative Therapy and High-throughput Drug Screening
Assessing Genotoxicity
-
Research • Cancer ResearchEvaluating In Vitro DNA Damage Using Comet Assay -
Research • GeneticsUse of Frozen Tissue in the Comet Assay for the Evaluation of DNA Damage -
Research • BiologyRapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae -
Research • NeuroscienceAssessing Cell Cycle Progression of Neural Stem and Progenitor Cells in the Mouse Developing Brain after Genotoxic Stress -
Research • BioengineeringAdvanced 3D Liver Models for In vitro Genotoxicity Testing Following Long-Term Nanomaterial Exposure -
Research • MedicineGene-environment Interaction Models to Unmask Susceptibility Mechanisms in Parkinson's Disease
Toxicology Testing
-
Research • BiologyHigh Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC) -
Research • BiologyExperimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory -
Research • Developmental BiologyHuman Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation -
Research • BiologyDetecting Estrogenic Ligands in Personal Care Products using a Yeast Estrogen Screen Optimized for the Undergraduate Teaching Laboratory -
Research • Developmental BiologyProtocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing
