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HTS091M ChemiSCREEN™ Human DP Prostanoid Receptor Membrane Preparation

Human DP GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ Human DP Prostanoid Receptor Membrane Preparation MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: Q13258 Entrez Gene Number: NM_000953.2
HTS091M
200 units  
Purchase on Sigma-Aldrich

Áttekintés

Replacement Information

Kulcsspecifikációk táblázata

SpeciesHost CellsProtein Target
Human Chem-1DP
Description
Catalogue NumberHTS091M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Human DP Prostanoid Receptor Membrane Preparation
OverviewFull-length human PTGDR cDNA encoding DP
Background InformationProstanoids are a series of arachidonic acid metabolites produced by the action of cyclooxygenase and subsequently by isomerases and synthases. Cells rapidly secrete prostanoids after synthesis, whereupon the prostanoids bind to a family of 8 GPCRs to exert their biological effects (Narumiya and FitzGerald, 2001). The prostaglandin PGD2 is produced by mast cells upon activation by allergens, and is present at high levels in allergic diseases. PGD2 binds to two receptors, DP and CRTH2. DP activates Gs to increase cAMP levels, and lack of DP results in reduced allergic response in animal models of bronchial asthma (Matsuoka et al., 2000). Chemicon's DP 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 DP interactions and its ligands. The membrane preparations exhibit a Kd of 10.39 nM for [3H]-PGD2. With 8 nM [3H]-PGD2, 5 µg/well and 10 µg/well DP Membrane Prep typically yield greater than 4-fold signal-to-background ratio.
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 DP GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPS binding.
Biological Information
Entrez Gene Number
Entrez Gene SummaryThe protein encoded by this gene is a G-protein-coupled receptor. It has been shown to function as a prostanoid DP receptor. The activity of this receptor is mainly mediated by G-S proteins that stimulate adenylate cyclase resulting in an elevation of intracellular cAMP and Ca2+. Knockout studies in mice suggest that the ligand of this receptor, prostaglandin D2 (PGD2), functions as a mast cell-derived mediator to trigger asthmatic responses.
Gene Symbol
  • PTGDR
  • DP
  • MGC49004
  • AS1
  • ASRT1
GPCR ClassA
Host CellsChem-1
Protein TargetDP
Target Sub-FamilyProstanoid
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q13258 # Receptor for prostaglandin D2 (PGD2). The activity of this receptor is mainly mediated by G(s) proteins that stimulate adenylate cyclase, resulting in an elevation of intracellular cAMP. A mobilization of calcium is also observed, but without formation of inositol 1,4,5-trisphosphate.
SIZE: 359 amino acids; 40271 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in retina and small intestine.
DISEASE:SwissProt: Q13258 # Genetic variations in PTGDR are associated with susceptibility to asthma-related traits type 1 (ASRT1) [MIM:607277]. Asthma-related traits include clinical symptoms of asthma, such as coughing, wheezing and dyspnea.
SIMILARITY: SwissProt: Q13258 ## 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, a GF/C 96-well filter plate 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: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [3H] PGD2 (Perkin Elmer#:NET-616 )
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 4-fold signal:background with 3H-labeled PGD2 at 8 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of DP membrane prep:
Product Usage Statements
10 µg/well 5 µg/well
Signal:Background 4.86 4.12
Specific Binding (cpm) 1345 885



Table 2. IC50 values for ligands obtained in a competition binding assay with DP membrane preparation.
IC50 (nM)
Prostaglanding D2 12.1
13,14-dihydro-15-keto prostaglandin D2 >10000
BW A868C 4.8


SPECIFICATIONS: 1 unit = 10 µg
Bmax for [3H] PGD2 binding: 5.3 pmol/mg protein;
Kd for [3H] PGD2 binding: ~10.39nM 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
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalógusszám GTIN
HTS091M 04053252624117

Documentation

ChemiSCREEN™ Human DP Prostanoid Receptor Membrane Preparation MSDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Human DP Prostanoid Receptor Membrane Preparation Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN DP PROSTANOID RECEPTOR - 2349734 2349734

References

Reference overviewPub Med ID
Genetic and pharmacological analysis of prostanoid receptor function
Narumiya, S and FitzGerald, G A
J Clin Invest, 108:25-30 (2001)  2001

11435452 11435452
Prostaglandin D2 as a mediator of allergic asthma
Matsuoka, T, et al
Science, 287:2013-7 (2000)  1999

10720327 10720327

Data Sheet

Title
CHEMISCREEN MEMBRANE PREPARATION RECOMBINANT HUMAN DP PROSTANOID RECEPTOR

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Kategóriák

Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations