HTS031M
ChemiSCREEN CRTH2 Membrane Preparation
Human CRTH2 / DP2 / GPR44 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Sign Into View Organizational & Contract Pricing
All Photos(2)
About This Item
UNSPSC Code:
41106514
eCl@ss:
32161000
NACRES:
NA.41
biological source
human
Quality Level
recombinant
expressed in Chem-1 cells
manufacturer/tradename
ChemiScreen
Chemicon®
technique(s)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBI accession no.
UniProt accession no.
shipped in
dry ice
General description
Chemoattractant 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.
Human CRTH2
Application
Radioligand binding assay, and GTPgammaS binding.
Biochem/physiol Actions
GPCR Class: A
Protein Target: CRTH2 / DP2 / GPR44
Target Sub-Family: Prostanoid
Packaging
1 mL
Quality
Signal:background and specific binding values obtained in a competition binding assay with CRTH2 membrane preparation with unlabeled Prostaglandin D2.
10 g/well
Signal:Background 12.2
Specific Binding (cpm) 4307
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 g
Bmax: 13.93 pmol/mg
Kd: 10 nM
10 g/well
Signal:Background 12.2
Specific Binding (cpm) 4307
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 g
Bmax: 13.93 pmol/mg
Kd: 10 nM
Specifications
Inucbation Conditions
Membranes 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)
Membranes 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)
Physical form
Liquid 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.
Storage and Stability
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Legal Information
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
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 Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
Already Own This Product?
Find documentation for the products that you have recently purchased in the Document Library.
K Nagata et al.
Journal of immunology (Baltimore, Md. : 1950), 162(3), 1278-1286 (1999-02-11)
The search for reliable marker molecules discriminating between human Th1 and Th2 cells identified a gene encoding a novel member of the G protein-coupled leukocyte chemoattractant receptor family, which is selectively expressed in Th2 but not Th1 lineage cells, thereby
Hiromi Sugimoto et al.
The Journal of pharmacology and experimental therapeutics, 305(1), 347-352 (2003-03-22)
Ramatroban (Baynas, BAY u3405), a thromboxane A(2) (TxA(2)) antagonist marketed for allergic rhinitis, has been shown to partially attenuate prostaglandin (PG)D(2)-induced bronchial hyperresponsiveness in humans, as well as reduce antigen-induced early- and late-phase inflammatory responses in mice, guinea pigs, and
T Matsuoka et al.
Science (New York, N.Y.), 287(5460), 2013-2017 (2000-03-17)
Allergic asthma is caused by the aberrant expansion in the lung of T helper cells that produce type 2 (TH2) cytokines and is characterized by infiltration of eosinophils and bronchial hyperreactivity. This disease is often triggered by mast cells activated
Michitaka Shichijo et al.
The Journal of pharmacology and experimental therapeutics, 307(2), 518-525 (2003-09-17)
We cloned, expressed, and characterized in vitro and in vivo the gene encoding the rat ortholog of chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2), a G protein-coupled receptor for prostaglandin D2 (PGD2). Quantitative reverse transcription-polymerase chain reaction analysis demonstrated
H Hirai et al.
The Journal of experimental medicine, 193(2), 255-261 (2001-02-24)
Prostaglandin (PG)D2, which has long been implicated in allergic diseases, is currently considered to elicit its biological actions through the DP receptor (DP). Involvement of DP in the formation of allergic asthma was recently demonstrated with DP-deficient mice. However, proinflammatory
Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.
Contact Technical Service