- AID 540340 - SAR analysis counterscreen of small molecule.
- PPSX Eurofins Pharma Services General Presentation - DiscoverX.
- Beta-Arrestin recruitment assay for the identification of.
- Measurement of β-Arrestin Recruitment for GPCR Targets.
- Table 1. [Comparison of various β-arrestin assay...]. - Assay.
- PathHunter® β-Arrestin Assays as a Universal Tool For Orphan GPCR.
- Why Study GPCR Arrestin Recruitment | Eurofins DiscoverX Blog.
- Cell-based high-throughput screening assay system for... - PubMed.
- Characterizing cannabinoid CB2 receptor ligands using DiscoveRx.
- PathHunter eXpress β-Arrestin Orphan GPCR Assays.
- DiscoverX Launches Toolbox Products for GPCR β-Arrestin Cell.
- PDF For Chemiluminescent Detection of Activated GPCRs.
- C3aR and C5aR1 act as key regulators of human and mouse β-cell function.
- Screening β-arrestin recruitment for the identification of natural.
AID 540340 - SAR analysis counterscreen of small molecule.
In the DiscoveRx (Fremont, CA) PathHunter βarr assay, a β-galactosidase enzyme is complemented upon βarr recruitment (Yin et al., 2009). Alternative experimental approaches are the GFP (YFP)-distribution assays (Fig. 1B), the CFP/YFP Förster resonance energy transfer assay technique (Fig. 1C), and the Tango assay format (Fig. 1D). PathHunter® eXpress GPR20 HEK 293 β-Arrestin Orphan GPCR Assay, DiscoverX. Access DiscoverX's extensive menu of functional, whole cell GPCR, Kinase, Interleukin, Pathway, NHR, and more assays without investing in a clonal cell line. Complete off-the-shelf assay solution; Pre-validated frozen cells and reagents; Fast and simple step-by-step. PathHunter β-arrestin Assay: DiscoverX: A β-galactosidase enzyme fragment complementation technology-based assay. β-Arrestin recruitment brings about reconstitution of an active β-galactosidase, which is split into a PK fragment fused to the GPCR and an enzyme acceptor protein fused to the β-arrestin.
PPSX Eurofins Pharma Services General Presentation - DiscoverX.
The new assays, which enable researchers to use the preferred cell models of their choice, include the HTRF GTP Gi binding kit, the industry's first TR-FRET based assay for GTP binding; the B-arr2 recruitment kit; and HTRF total kits for B-Arrestin 1, B-Arrestin 2 and AP2. These assays will help scientists continue to better understand the. Assays monitoring beta-arrestin related pathways: β-arrestin recruitment: PathHunter (DiscoverX) Split β-galactosidase enzyme fragments fused to GPCR and β-arrestin. Arrestin recruitment results in functional complementation and a luminescent enzyme product when substrate is present: Upstream event is monitored; High-throughput amenable.
Beta-Arrestin recruitment assay for the identification of.
These assays generally require a cell line coexpressing a fluorophore-tagged GPCR of interest and a luciferase-tagged β-arrestin. Upon GPCR-β-arrestin interaction, a BRET (bioluminescence resonance energy transfer) signaling is generated by proximity of the two tags. Reporter gene-based assays for β-arrestin activity are also available. DiscoveRx Corporation 42501 Albrae Street Fremont, CA 94538 USA tel | 1.510.979.1415 x104 For some products/cell lines, certain 3rdparty gene specific patents may be required to use the cell line. It is the purchaser's responsibility to determine if such patents or other intellectual property rights are required. 4. In both cases, the assays employ the industry-validated Dx enzyme fragment complementation system which consists of a split beta-galactodase enzyme into two non-functional fragments which we call enzyme donor(ED) or ProLink and enzyme acceptor(EA). The ED is a small peptide where the EA is the bulk of the beta-gal enzyme.
Measurement of β-Arrestin Recruitment for GPCR Targets.
However, for the majority of orphan receptors studied, measurement of β-arrestin recruitment did not lead to the identification of cognate ligands from our screening sets. β-Arrestin recruitment represents a robust GPCR screening technology, and ligand-biased signaling is emerging as a therapeutically exploitable feature of GPCR biology. MultiScreen β-Arrestin Sensor Technology enables researchers to: Assess GPCRs in their native form for true pharmacology. Assay endogenously expressed GPCRs for more relevant data. Characterize orphan GPCRs to expand your target pool. Use a single cell line for multiple GPCR assays for accelerated drug development. Β-arrestin Angiotensin AT1R Assay. β-arrestin Vasopressin V2R Assay. β-arrestin GLP-1R assay. β-arrestin Mu Opioid assay. β-arrestin β2AR assay. β-arrestin D1 Dopamine assay. Detect agonist bias in living cells. Measure kinetic cellular responses. Monitor Arrestin and G-limb signaling using the same modality.
Table 1. [Comparison of various β-arrestin assay...]. - Assay.
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PathHunter® β-Arrestin Assays as a Universal Tool For Orphan GPCR.
Nov 20, 2017 · All four assays involve the expression of the β-arrestin as a fusion protein with another protein or fragment, while the PathHunter, Tango and LinkLight assays require fusion of the GPCR to another peptide or protein moiety as well. Table 1 compares the principles of these technologies as well as their advantages and limitations.. HitHunter® cAMP assays are competitive immunoassays that utilize EFC. Beta arrestin assays can also be used to monitor cAMP levels by TRFRET >390 Assays are offered in Agonist, Antagonist, PAM and NAM modes: Primary screening at one or more concentration. Dose response format at 10 concentration in duplicate. Assays are also available for HTS.
Why Study GPCR Arrestin Recruitment | Eurofins DiscoverX Blog.
DiscoverX beta-arrestin reporter cell assays indicated that commercially available selective agonists for C3aR (sc-214644) and C5aR1 (65121-ANA) induced concentration-dependent activation of C3aR and C5aR1, with half-maximal activation at 804 nM sc-214644 and 292 nM 65121-ANA, respectively (Fig. 3 a, b). Beta-Arrestin recruitment assays provide a generic assay platform for drug discovery on G-protein-coupled receptors (GPCRs). The PathHunter assay technology developed by DiscoveRx (Fremont, CA) uses enzyme fragment complementation of beta-galactosidase to measure receptor-beta-arrestin proximity by chemiluminescence. Contact Information DiscoveRx Corporation World Wide Headquarters 42501 Albrae Street Fremont CA 94538 United States t 5109791415 f 5109791650 toll-free 8664484864 KINOMEscan….
Cell-based high-throughput screening assay system for... - PubMed.
DiscoveRx Corporation 42501 Albrae Street Fremont, CA 94538 USA tel | 1.510.979.1415 x104 For some products/cell lines, certain 3rdparty gene specific patents may be required to use the cell line. It is the purchaser's responsibility to determine if such patents or other intellectual property rights are required. 4. Here we present the PathHunter ® Arrestin assay as a facile screening approach to orphan GPCR drug discovery. In this system, a small peptide is fused to the GPCR target, and the complementing fragment is fused to β-Arrestin2. Upon activation, the GPCR binds to arrestin forcing complementation of the enzyme fragments.
Characterizing cannabinoid CB2 receptor ligands using DiscoveRx.
.. Jan 14, 2019 · The PathHunter β-arrestin assay (Discoverx) was carried out with the CHO-β 2-β-arr cell line expressing the Prolink-tagged human β 2-adrenoceptors and the β-arrestin-2-β-galactosidase enzyme.
PathHunter eXpress β-Arrestin Orphan GPCR Assays.
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DiscoverX Launches Toolbox Products for GPCR β-Arrestin Cell.
Jun 28, 2016 · FREMONT, Calf., June 28, 2016 /PRNewswire/ -- DiscoverX Corporation, the leading supplier of innovative cell-based assays and services for drug discovery and development, today announced the. Nov 20, 2017 · The PathHunter GPCR β-arrestin assay is based on proprietary Enzyme Fragment Complementation technology from DiscoverX. PathHunter β-arrestin GPCR cells are engineered to co-express the ProLink (PK) tagged GPCR and the Enzyme Acceptor (EA) tagged β-arrestin.
PDF For Chemiluminescent Detection of Activated GPCRs.
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C3aR and C5aR1 act as key regulators of human and mouse β-cell function.
. Arrestin recruitment occurs independent of G-protein coupling, these assays provide a direct, universal platform for measuring receptor activation. Figure 1. PathHunter β-Arrestin Assay Principle. Activation of the ProLink-tagged GPCR results in β-Arrestin recruitment and formation of a functional enzyme capable of hydrolyzing. Mar 01, 2010 · In this assay, the GPCR is fused at the C-terminus to a mutant peptide fragment of the β-galactosidase (Prolink™ or enzyme donor) and the β-arrestin is tagged with the corresponding deletion mutant of β-galactosidase (enzyme acceptor) which is provided by DiscoveRx, engineered into host cells [31, 32].
Screening β-arrestin recruitment for the identification of natural.
Arrestin recruitment occurs independent of G-protein coupling, these assays provide a direct, universal platform for measuring receptor activation. Figure 1. PathHunter® β-Arrestin Assay Principle. Activation of the ProLink-tagged GPCR results in β-Arrestin recruitment and formation of a functional enzyme capable of hydrolyzing. The HTRF cAMP assay (Cisbio Bioassays) was conducted on compound-stimulated HEK293 cells expressing a low level of an endogenous β 2-adrenoceptor. The PathHunter β-arrestin assay (Discoverx) was conducted on compound-stimulated CHO-β 2-β-arr cells. Both of these cell lines have no background expression of other β-adrenoceptor isoforms. Assays to Measure Ligand Bias The development of biased ligands requires tools to measure signaling via different pathways plus methods for analyzing results in order to quantify bias. Assays for β-arrestin recruitment and signaling via cAMP modulation or calcium mobilization are needed to define compound activity in both potency and efficacy.
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