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HTS103M ChemiSCREEN™ Human D3 Dopamine Receptor Membrane Preparation

Human D3 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P35462 Entrez Gene Number: NM_000796.3, NM_033663.3
HTS103M
200 units  
Purchase on Sigma-Aldrich

概述

Replacement Information

重要规格表

SpeciesHost CellsProtein Target
Human Chem-1D3
Description
Catalogue NumberHTS103M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Human D3 Dopamine Receptor Membrane Preparation
OverviewHuman D3 Receptor
Background InformationDopamine is a catecholamine neurotransmitter that functions in the CNS to control locomotor, cognitive, emotional and neurendocrine processes, and in the periphery to modulate cardiovascular, renal and gastrointestinal processes. The biological activities of dopamine are mediated by a family of five GPCRs. The D1 and D5 subtypes couple to Gs to increase intracellular cAMP, whereas the D2, D3 and D4 subtypes couple to Gi to reduce cAMP (Missale et al., 1998). The dopamine D3 receptor is expressed in phylogenetically older regions of the brain, suggesting that it plays a role in cognitive and emotional functions. It has been the target of potential drugs for psychotic disorders and Parkinson's disease. Chemicon's D3 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 D3 interactions and its ligands. The membrane preparations exhibit a Kd of 0.1 nM for [125I]-Iodospiperone. With 5.0ug/well D3 membrane prep and 0.1 nM [125I]-Iodospiperone, a greater than 4-fold signal-to-background ratio was obtained.
References
Product Information
FormatMembranes
PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 4-fold signal:background with 125I-labeled Iodospiperone at 0.1 nM.

Liquid in packaging buffer: 50 mM Tris, pH 7.4, 10% glycerol with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Quality LevelMQ100
Applications
ApplicationHuman D3 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPγS binding
Biological Information
Entrez Gene Number
Entrez Gene SummaryThis gene encodes the D3 subtype of the dopamine receptor. The D3 subtype inhibits adenylyl cyclase through inhibitory G-proteins. This receptor is expressed in phylogenetically older regions of the brain, suggesting that this receptor plays a role in cognitive and emotional functions. It is a target for drugs which treat schizophrenia, drug addiction, and Parkinson disease. Alternative splicing of this gene results in multiple transcript variants that would encode different isoforms, although some variants may be subject to nonsense-mediated decay (NMD).
Gene Symbol
  • DRD3
  • FET1
  • MGC149204
  • D3DR
  • ETM1
  • D
  • MGC149205
GPCR ClassA
Host CellsChem-1
Protein TargetD3
Target Sub-FamilyDopamine
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P35462 # This is one of the five types (D1 to D5) of receptors for dopamine. The activity of this receptor is mediated by G proteins which inhibit adenylyl cyclase.
SIZE: 400 amino acids; 44225 Da
SUBUNIT: Interacts with CLIC6 (By similarity).
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Brain.
DISEASE: SwissProt: P35462 # Genetic variation in DRD3 may be associated with susceptibility to hereditary essential tremor 1 (ETM1) [MIM:190300]. ETM1 is the most common movement disorder. The main feature is postural tremor of the arms. Head, legs, trunk, voice, jaw, and facial muscles also may be involved. The condition can be aggravated by emotions, hunger, fatigue and temperature extremes, and may cause a functional disability or even incapacitation. Inheritance is autosomal dominant.
SIMILARITY: SwissProt: P35462 ## 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 Tris, pH 7.4. 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 Tris, pH 7.4, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] -Iodospiperone(Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
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 varying amounts of D3 Receptor membrane prep.
Product Usage Statements
10 µg/well 5 µg/well
Signal:Background 4.88 5.93
Specific Binding (cpm) 1853.7 1861.3



SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 0.32 pmol/mg
Kd: 0.1 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
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
产品目录编号 GTIN
HTS103M 04053252620614

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