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HTS031M ChemiSCREEN™ CRTH2 Membrane Preparation

Human CRTH2 / DP2 / GPR44 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ CRTH2 Membrane Preparation: MSDS (material safety data sheet) o SDS, certificato d’analisi (CoA) e certificato di qualità (CoQ), dossier, brochure e altri documenti disponibili.
Host Cell: Chem-1 Uni Prot Number: Q9Y5Y4 Entrez Gene Number: NM_004778.2
HTS031M
200 units  1 mL
Purchase on Sigma-Aldrich

Panoramica

Replacement Information

Tabella delle specifiche principali

SpeciesHost CellsProtein Target
Human Chem-1CRTH2 / DP2 / GPR44
Description
Catalogue NumberHTS031M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ CRTH2 Membrane Preparation
OverviewHuman CRTH2
Background InformationChemoattractant receptor homologous molecule expressed on Th2 cells (CRTH2) is a GPCR related to FPR-like chemoattractant receptors that is highly expressed on Th2 cells, eosinophils, basophils, and type 2 cytotoxic T cells (Nagata et al., 1999). The most potent ligand for CRTH2 is the prostaglandin PGD2, a major mast cell-derived allergic mediator (Hirai et al., 2001). Another GPCR, DP, is activated by PGD2, and the relative contributions of DP and CRTH2 to PGD2-mediated allergic disease are being elucidated. A small molecule antagonist of CRTH2, ramatroban, attenuates PGD2-mediated bronchial hyperresponsiveness and antigen-induced inflammation (Sugimoto et al., 2003; Shichijo et al., 2003).Chemicon's CRTH2 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of PGD2/CRTH2 interactions. The membrane preparations exhibit a Kd of 10 nM for [3H]-Prostaglandin D2. With 2.5 μg/well CRTH2 Membrane Prep and 10 nM [3H]-Prostaglandin D2, a greater than 5-fold signal-to-background ratio is obtained.
References
Product Information
FormatMembranes
PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman CRTH2 / DP2 / GPR44 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding.
Biological Information
Entrez Gene Number
Entrez Gene SummaryG protein-coupled receptors (GPCRs), such as GPR44, are integral membrane proteins containing 7 putative transmembrane domains (TMs). These proteins mediate signals to the interior of the cell via activation of heterotrimeric G proteins that in turn activate various effector proteins, ultimately resulting in a physiologic response.[supplied by OMIM]
Gene Symbol
  • GPR44
  • DL1R
  • CRTH2
  • CD294
GPCR ClassA
Host CellsChem-1
Protein TargetCRTH2 / DP2 / GPR44
Target Sub-FamilyProstanoid
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q9Y5Y4 # Orphan receptor.
SIZE: 395 amino acids; 43240 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: Q9Y5Y4 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsMembranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.
Binding buffer: 50mM HEPES, 10mM MnCl2, 1mM EDTA, 0.2% BSA, pH to 7.0 and filter.
Wash buffer: 50mM HEPES, 0.5M NaCl, pH to 7.4 and filter.
Radioligand: [3H] Prostaglandin D2 (Amersham#: TRK734)
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with CRTH2 membrane preparation with unlabeled Prostaglandin D2.
Product Usage Statements
10 µg/well
Signal:Background 12.2
Specific Binding (cpm) 4307

Table 2. IC50 values for ligands obtained in a competition binding assay with CRTH2 membrane preparation.
IC50 (nM)
Prostaglanding D2 4.5
13,14-dihydro-15-keto prostaglandin D2 14.7
BW A868C 7429


SPECIFICATIONS: 1 unit = 10 µg
Bmax: 13.93 pmol/mg
Kd: 10 nM
Usage Statement
  • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Material Package1 mL
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Numero di catalogo GTIN
HTS031M 04053252581724

Documentation

Protocols

Title
Assembly of a Competition Binding Curve

ChemiSCREEN™ CRTH2 Membrane Preparation MSDS

Titolo

Scheda di sicurezza (MSDS) 

Riferimenti bibliografici

Panoramica dei riferimenti bibliograficiCodice d'identificazione nel Pub Med
An orally bioavailable small molecule antagonist of CRTH2, ramatroban (BAY u3405), inhibits prostaglandin D2-induced eosinophil migration in vitro
Sugimoto, Hiromi, et al
J Pharmacol Exp Ther, 305:347-52 (2003)  2003

12649388 12649388
Chemoattractant receptor-homologous molecule expressed on Th2 cells activation in vivo increases blood leukocyte counts and its blockade abrogates 13,14-dihydro-15-keto-prostaglandin D2-induced eosinophilia in rats
Shichijo, Michitaka, et al
J Pharmacol Exp Ther, 307:518-25 (2003)  2003

12975488 12975488
Prostaglandin D2 selectively induces chemotaxis in T helper type 2 cells, eosinophils, and basophils via seven-transmembrane receptor CRTH2
Hirai, H, et al
J Exp Med, 193:255-61 (2001)  2001

11208866 11208866
Prostaglandin D2 as a mediator of allergic asthma
Matsuoka, T, et al
Science, 287:2013-7 (2000)  1999

10720327 10720327
Selective expression of a novel surface molecule by human Th2 cells in vivo
Nagata, K, et al
J Immunol, 162:1278-86 (1999)  1998

9973380 9973380

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Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations