標(biāo)題: Titlebook: G Protein-Coupled Receptor Screening Assays; Methods and Protocol Sofia Aires M. Martins,Duarte Miguel F. Prazeres Book 2021Latest edition [打印本頁] 作者: estrange 時間: 2025-3-21 19:04
書目名稱G Protein-Coupled Receptor Screening Assays影響因子(影響力)
作者: intrude 時間: 2025-3-21 21:03
Detection of GPCR mRNA Expression in Primary Cells Via qPCR, Microarrays, and RNA-Sequencing,ss GPCR mRNAs. Beginning from the isolation of cDNA and preparation of mRNA, we provide protocols for designing and testing qPCR primers, assaying mRNA expression using qPCR and high-throughput analysis of GPCR mRNA expression via TaqMan qPCR-based, GPCR-selective arrays. We also provide a workflow 作者: Coeval 時間: 2025-3-22 00:35 作者: 要塞 時間: 2025-3-22 08:26
Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor,ties of GPCR and mechanism of activity, these proteins should be isolated in significant (milligram) quantities, in a pure, homogenous, and stable form. Here we describe the expression and purification of type II human cannabinoid receptor CB., a class A GPCR, in two different types of expression ho作者: 統(tǒng)治人類 時間: 2025-3-22 11:55 作者: 拒絕 時間: 2025-3-22 16:16
Immobilization of Olfactory Receptors Carried by Nanosomes onto a Gold Sensor Surface,ach having specific odorant ligands. ORs could be used as sensing elements of highly specific and sensitive bioelectronic hybrid devices such as bioelectronic noses. After optimized immobilization onto the device, natural ORs provide molecular recognition of various odors with their intrinsic sensit作者: 拒絕 時間: 2025-3-22 18:59
Screening Methods for Cell-Free Synthesized GPCR/Nanoparticle Samples,ssembled nanodiscs allows the co-translational insertion of membrane proteins into tailored lipid bilayers in the absence of any artificial hydrophobic compounds. This strategy is particularly interesting for detergent sensitive or otherwise critical membrane proteins such as G-protein-coupled recep作者: 創(chuàng)新 時間: 2025-3-22 23:52
Fluorescence Anisotropy-Based Assay for Characterization of Ligand Binding Dynamics to GPCRs: The Cver, only a few of the assays enable studying wild-type receptors and monitor the ligand binding in real time. One of the approaches that is inherently suitable for this purpose is the fluorescence anisotropy (FA) assay. In the FA assay, the change of ligand’s rotational freedom connected with its b作者: Semblance 時間: 2025-3-23 04:37
Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Tecctivation in intact cells. This technology can be easily adopted to various plate reader devices and microtiter plate formats. Due to the high sensitivity of these BRET-based receptor biosensors and their ability to quantify simultaneously receptor activation/de-activation kinetics as well as compou作者: HUMP 時間: 2025-3-23 08:01 作者: ablate 時間: 2025-3-23 10:44
Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity,nable dynamic measurements. Moreover, FRET biosensors are ideally suited for the analysis of single living cells. The FRET biosensors described in this manuscript are entirely genetically encoded by plasmids. Here, protocols for employing FRET-based biosensors to detect G protein activity upon GPCR 作者: Certainty 時間: 2025-3-23 17:37
cAMP Biosensor Assay Using BacMam Expression System: Studying the Downstream Signaling of LH/hCG Rehe biological activity of various GPCR-specific agents, including protein hormones. cAMP biosensors based on detection of F?rster-type resonance energy transfer (FRET) offer unique advantages including the ratiometric nature of measurement, adjustable affinity toward detected molecule, capability of作者: 微粒 時間: 2025-3-23 18:42
FLIPR Calcium Mobilization Assays in GPCR Drug Discovery,fluorescence-based method (FLIPR Calcium 4 Assay) developed by Molecular Devices for a FlexStation and routinely used in our laboratory for detecting intracellular calcium changes. The assay is designed to study calcium mobilization induced by majority of GPCRs and calcium channels and allows for si作者: 禁止,切斷 時間: 2025-3-24 01:00 作者: 羅盤 時間: 2025-3-24 02:35
Split-Tobacco Etch Virus (Split-TEV) Method in G Protein-Coupled Receptor Interacting Proteins,(TEV) protease in two fragments, where each fragment is fused to the candidate proteins predicted to interact. If there is indeed an interaction between both proteins, TEV protease reconstitutes its proteolytic activity and this activity is used to induce the expression of some reporter genes. Howev作者: CUR 時間: 2025-3-24 08:39 作者: sleep-spindles 時間: 2025-3-24 11:33 作者: 遺忘 時間: 2025-3-24 15:10
Gradient Tracking by Yeast GPCRs in a Microfluidics Chamber,n provide precise control of the time, dose, and orientation of a stimulus, while simultaneously capturing quantitative single-cell data. The approach is particularly powerful when combined with the genetically tractable yeast model organism. The GPCR pathway in yeast is structurally conserved and f作者: 人充滿活力 時間: 2025-3-24 20:32
Monitoring Intracellular Calcium in Response to GPCR Activation: Comparison Between Microtiter Platr speeds and lower costs, while at the same time mimicking the local chemical concentrations and force fields of the cellular in vivo environment. In this chapter we introduce a microfluidic structure with hydrodynamic cell traps and a culture volume in the nanoliter range (50?nL), to quantitatively作者: 他去就結(jié)束 時間: 2025-3-25 00:36 作者: Cumbersome 時間: 2025-3-25 04:17 作者: STYX 時間: 2025-3-25 11:27 作者: 額外的事 時間: 2025-3-25 13:58 作者: 過剩 時間: 2025-3-25 17:03
Split-Tobacco Etch Virus (Split-TEV) Method in G Protein-Coupled Receptor Interacting Proteins,of the Split-TEV method developed with the aim to increase the specificity in the study of G protein-coupled receptor (GPCR) interacting proteins. This approach for monitoring interactions between GPCRs is an easy and robust assay and offers good perspectives in drug discovery.作者: Contort 時間: 2025-3-25 23:54
,NanoLuc-Based Methods to Measure β-Arrestin2 Recruitment to G Protein-Coupled Receptors, enzyme-fragment complementation in real-time on living cells with reasonable high throughput. In addition to the detection of agonism, both assay formats can be used to qualitatively evaluate the binding kinetics of antihistamines on the human histamine H. receptor.作者: Limited 時間: 2025-3-26 00:57
1064-3745 expertsThis fully updated edition targets not only those assays directly involved in the discovery of GPCR-active compounds but also those involved in cell-based experiments designed to study physiological responses. Whether coming from academia or industry, or being an experienced researcher or a 作者: seruting 時間: 2025-3-26 07:36 作者: 比目魚 時間: 2025-3-26 10:26
Studium, Ausbildung und Fortbildung,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.作者: 蚊帳 時間: 2025-3-26 15:24 作者: 萬神殿 時間: 2025-3-26 16:49 作者: 發(fā)起 時間: 2025-3-26 22:38
Frank Brettschneider,Markus Rettich means of BacMam transduction system. As a practical challenge, we address the applicability of the designed assay for screening of biological activity of human hormones, including human chorionic gonadotropin (hCG) bearing different posttranslational modifications.作者: 外觀 時間: 2025-3-27 04:46 作者: 使成整體 時間: 2025-3-27 08:17
cAMP Biosensor Assay Using BacMam Expression System: Studying the Downstream Signaling of LH/hCG Re means of BacMam transduction system. As a practical challenge, we address the applicability of the designed assay for screening of biological activity of human hormones, including human chorionic gonadotropin (hCG) bearing different posttranslational modifications.作者: excrete 時間: 2025-3-27 10:58
Book 2021Latest editionased experiments designed to study physiological responses. Whether coming from academia or industry, or being an experienced researcher or a newcomer to the field, the reader will find accessible methods and protocols that cover the latest developments on receptor purification, molecular biology, r作者: PAD416 時間: 2025-3-27 16:47
https://doi.org/10.1007/978-1-4899-3468-0sts: in . and in mammalian suspension cell culture Expi293. Our method allows preparation of milligram quantities of the purified receptors suitable for a wide array of downstream applications including high-resolution structural studies and functional assays.作者: 業(yè)余愛好者 時間: 2025-3-27 19:10 作者: 受傷 時間: 2025-3-27 21:56 作者: PANIC 時間: 2025-3-28 03:15
https://doi.org/10.1007/978-3-322-91586-3 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).作者: Oratory 時間: 2025-3-28 08:44
Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor,sts: in . and in mammalian suspension cell culture Expi293. Our method allows preparation of milligram quantities of the purified receptors suitable for a wide array of downstream applications including high-resolution structural studies and functional assays.作者: 下船 時間: 2025-3-28 13:21
Bioluminescence in G Protein-Coupled Receptors Drug Screening Using Nanoluciferase and Halo-Tag Tecnd efficacy and potency, these optical tools provide the most direct and unbiased approach to monitor GPCR activity in a high-throughput-compatible assay format, representing a novel promising tool for the discovery of potential GPCR therapeutics.作者: generic 時間: 2025-3-28 15:28 作者: 擁護(hù)者 時間: 2025-3-28 20:10
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).作者: Flustered 時間: 2025-3-29 02:12
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.作者: Mettle 時間: 2025-3-29 03:08
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.作者: 古文字學(xué) 時間: 2025-3-29 08:47
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.作者: Mawkish 時間: 2025-3-29 14:42 作者: NADIR 時間: 2025-3-29 17:01 作者: Wordlist 時間: 2025-3-29 22:04 作者: 冷淡周邊 時間: 2025-3-30 03:03
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作者: Graphite 時間: 2025-3-30 04:23
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作者: phytochemicals 時間: 2025-3-30 11:01 作者: 博識 時間: 2025-3-30 15:33
Luciferase Complementation Approaches to Measure GPCR Signaling Kinetics and Bias,r kinetic properties. Recent developments in complementation approaches, based on a small bright shrimp luciferase Nanoluc, provide a new route to kinetic analysis of GPCR signaling in living cells that is amenable to the throughput required for compound profiling. In the NanoBiT luciferase compleme作者: 護(hù)航艦 時間: 2025-3-30 20:16
https://doi.org/10.1007/978-3-8348-8201-1 state and upon receptor agonism, and the evaluation of the biological functions of the GPCR in appropriate cell lines. The lack of standardized methodologies to study lipid rafts, in general, and of the workings of GPCRs in lipid rafts, in particular, and the inescapable drawbacks of current method作者: 描述 時間: 2025-3-30 21:12
https://doi.org/10.1007/978-3-8348-1976-5maximize sensitivity while minimizing noise. These methods may also be applicable for assessing the expression of members of families of other low abundance genes via high-throughput analyses of mRNAs, followed by independent confirmation and validation of results via qPCR.作者: CBC471 時間: 2025-3-31 02:14 作者: Obligatory 時間: 2025-3-31 08:12 作者: ENACT 時間: 2025-3-31 12:42 作者: Migratory 時間: 2025-3-31 13:44
https://doi.org/10.1007/978-3-322-91961-8detecting G protein activation monitor the generation of second messengers ([Ca.]., cAMP, IP.) or active G proteins (with GTP-g-S for instance). While these approaches have proven sensitive and robust, they are not suited for the detection of a single GPCR-G protein interaction. Here, we present in 作者: CAPE 時間: 2025-3-31 18:46 作者: 關(guān)心 時間: 2025-4-1 00:49
Book 2021Latest editionown pitfalls.?.Authoritative and up-to-date, .G Protein-Coupled Receptor Screening Assays: Methods and Protocols, Second Edition. aims to provide the tools necessary to contribute to the advancement of GPCR research and discovery and ultimately lead to the availability of innovative and more efficie作者: 極大痛苦 時間: 2025-4-1 02:30 作者: 盡責(zé) 時間: 2025-4-1 09:20
978-1-0716-1223-1Springer Science+Business Media, LLC, part of Springer Nature 2021
