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ChemiScreen™ Membrane Prep Hu Recombinant CRTH2

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货号：HTS031M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 CRTH2 / DP2 / GPR44
Description
Catalogue Number HTS031M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ CRTH2 Membrane Preparation
Overview Human CRTH2
Background Information 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.
References
Product Information
Format Membranes
Presentation 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.
Applications
Application Human CRTH2 / DP2 / GPR44 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay, and GTPgammaS binding.
Biological Information
Entrez Gene Number
Entrez Gene Summary G 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 Class A
Host Cells Chem-1
Protein Target CRTH2 / DP2 / GPR44
Target Sub-Family Prostanoid
UniProt Number
UniProt Summary FUNCTION: 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 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)
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. 2
Signal:background and specific binding values obtained in a competition binding assay with CRTH2 membrane preparation with unlabeled Prostaglandin D.
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 Conditions Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Material Package 1 mL
Transport Information
Supplemental Information
Specifications

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