Titlebook: G Protein-Coupled Receptor Screening Assays; Methods and Protocol Sofia Aires M. Martins,Duarte Miguel F. Prazeres Book 2021Latest edition

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Monitoring Intracellular Calcium in Response to GPCR Activation: Comparison Between Microtiter Plat evaluate the transient calcium response of the endogenous Muscarinic type 1 receptor (M1) in HEK 293?T cells. The microfluidic fabrication protocol is described as well as a methodology to monitor the cell response in real time, after stimulation with M1 agonists (e.g., carbachol) and antagonists (e.g., .irenzepine).

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Screening for Serotonin Receptor 4 Agonists Using a GPCR-Based Sensor in Yeast,ally, the HTR.-based screen couples activation of 5-HTR. on the yeast cell surface to luciferase reporter expression. The HTR.-based screen has a throughput of one compound per second allowing the screening of more than a thousand compounds per day.

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Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity,s manuscript are entirely genetically encoded by plasmids. Here, protocols for employing FRET-based biosensors to detect G protein activity upon GPCR activation are reported. The protocols include details on the isolation of plasmids, transfection, generation of stable cell lines with the FRET biosensors, FRET ratio imaging, and data analysis.

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FLIPR Calcium Mobilization Assays in GPCR Drug Discovery,intracellular calcium changes. The assay is designed to study calcium mobilization induced by majority of GPCRs and calcium channels and allows for simultaneous concentration-dependent analysis of several receptor agonists and antagonists, useful in receptor characterization and drug discovery projects.

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Screening Methods for Cell-Free Synthesized GPCR/Nanoparticle Samples,cle membranes are excellent tools for a variety of applications such as ligand screening, engineering or even structural characterization. In this chapter, we provide protocols for the reaction set-up and efficient cell-free production of functionally folded GPCRs reaching μM concentrations in the f

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Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs: The Crmat limiting the flexibility of data analysis. To solve this problem, we propose the use of the data curation software . (.), which integrates experimental data with metadata in a Minimum Information for Data Analysis in Systems Biology (MIDAS) format. . enables data export to different software pa