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HTS129M ChemiScreen™ D5 Dopamine Receptor Membrane Preparation

Human D5 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

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Host Cell: Chem-1 Uni Prot Number: P21918 Entrez Gene Number: NM_000798.3
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HTS129M
200 units  
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      Übersicht

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1D5
      Description
      Catalogue NumberHTS129M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ D5 Dopamine Receptor Membrane Preparation
      OverviewFull-length human DRD5 cDNA encoding D5
      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 hypertensive phenotype of mice with a targeted deletion of D5 indicates that D5 regulates central control of sympathetic vascular tone (Hollon et al., 2002). In addition, D5 modulates locomotion and corticostriatal long-term depression (Centonze et al., 2003). Chemicon's cloned human D5-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant D5 expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between D5 and its ligands. The membrane preparations exhibit a Kd of 2.25 nM for [3H]-SCH 23390. With 0.5 nM [3H]-SCH 23390, 10 µg/well D5 Membrane Prep yields greater than 3-fold signal-to-background ratio.
      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 1 mg/mL in 1 mL packaging buffer, rapidly frozen, and stored at -80%deg;C.
      Quality LevelMQ100
      Applications
      ApplicationHuman D5 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 D5 subtype of the dopamine receptor. The D5 subtype is a G-protein coupled receptor which stimulates adenylyl cyclase. This receptor is expressed in neurons in the limbic regions of the brain. It has a 10-fold higher affinity for dopamine than the D1 subtype. Pseudogenes related to this gene reside on chromosomes 1 and 2.
      Gene Symbol
      • DRD5
      • DRD1B
      • DBDR
      • DRD1L2
      • D
      • MGC10601
      GPCR ClassA
      Host CellsChem-1
      Protein TargetD5
      Target Sub-FamilyDopamine
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P21918 # 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 activate adenylyl cyclase.
      SIZE: 477 amino acids; 52951 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Neuron-specific, localized primarily within limbic regions of the brain.
      DISEASE: SwissProt: P21918 # Defects in DRD5 may be a cause of schizophrenia, but no proof has yet been found. & Defects in DRD5 are a cause of blepharospasm [MIM:606798]. It is a primary focal dystonia affecting the orbicularis oculi muscles, usually beginning in middle age. Initial symptoms include eye irritation and frequent blinking, progressing to involuntary spasms of eyelid closure. In severe cases, this can lead to functional blindness.
      SIMILARITY: SwissProt: P21918 ## 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]-SCH 23390. (Perkin Elmer#:NET-930 )
      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 3-fold signal:background with 3H labeled SCH 23390 at 0.5 nM
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of D5 membrane prep:
      Product Usage Statements
      10 µg/well
      Signal:Background 4.06
      Specific Binding (cpm) 1952


      SPECIFICATIONS: 1 unit = 10 µg
      Bmax for [3H] SCH 23390 binding: 11.42 pmol/mg protein;
      Kd for [3H] SCH 23390 binding: ~ 2.25 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
      Global Trade Item Number
      Bestellnummer GTIN
      HTS129M 04053252731112