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HTS057M ChemiScreen™ Recombinant Human PRP/GPR10 Prolactin-Releasing Peptide Receptor Membrane Preparation

Human PRP / PrRP / GPR10 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Host Cell: Chem-1 Uni Prot Number: P49683 Entrez Gene Number: NM_004248.2
HTS057M
200 units  1 mL, 1 mg/mL
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      Aperçu

      Tableau de caractéristiques principal

      SpeciesHost CellsProtein Target
      Human Chem-1PRP / PrRP / GPR10
      Description
      Catalogue NumberHTS057M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ Recombinant Human PRP/GPR10 Prolactin-Releasing Peptide Receptor Membrane Preparation
      OverviewHuman Full-length human GPR10 cDNA encoding PRP
      Background InformationPRP, also known as GPR10 or hGR3, is a Gq-coupled receptor for prolactin-releasing peptide that is expressed in the pituitary (Hinuma et al., 1998). Genetic studies in rodents indicate that lack of GPR10 leads to hyperphagia, obesity and dyslipidemia (Gu et al., 2004; Watanabe et al., 2005). In humans, genetic variations in GPR10 are associated with lowered blood pressure (Bhattacharyya et al., 2003). Chemicon's GPR10/PRP 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 GPR10/PRP interactions with prolactin-releasing peptide. The membrane preparations exhibit a Kd of 0.59-0.66 nM for [125I]-PRP-20. With 5 µg/well PRP Membrane Prep and 0.25 nM [125I]-PRP-20, a greater than 30-fold signal-to-background ratio was obtained.
      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 30-fold signal:background with 125I-labeled PRP-20 at 0.25 nM.

      Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
      Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Quality LevelMQ100
      Applications
      ApplicationHuman PRP / PrRP / GPR10 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 SummaryPRLHR is a 7-transmembrane domain receptor for prolactin-releasing hormone (PRLH; MIM 602663) that is highly expressed in anterior pituitary (Ozawa et al., 2002 [PubMed 11923475]).[supplied by OMIM]
      Gene Symbol
      • PRLHR
      • GR3
      • GPR10
      • MGC126539
      • PrRPR
      • hGR3
      • MGC126541
      GPCR ClassA
      Host CellsChem-1
      Protein TargetPRP / PrRP / GPR10
      Target Sub-FamilyProlactin-releasing peptide
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P49683 # Receptor for prolactin-releasing peptide (PrRP). Implicated in lactation, regulation of food intake and pain-signal processing.
      SIZE: 370 amino acids; 41107 Da
      SUBUNIT: Interacts through its C-terminal region with the PDZ domain-containing proteins GRIP1, GRIP2 and PICK1. Interacts with PDZ domains 4 and 5 of GRIP1 and with the PDZ domain of PICK1.
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Only detected in the pituitary gland and in all cell types of pituitary adenomas.
      SIMILARITY: SwissProt: P49683 ## 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: [125I] PRP-20 (Perkin Elmer # NEX384)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Signal:background and specific binding values obtained in a competition binding assay with varying amounts of PRP membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 46.0 51.6
      Specific Binding (cpm) 15756 14148


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax: 8.9 pmol/mg
      Kd: 0.62 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
      Material Package1 mL, 1 mg/mL
      Global Trade Item Number
      Référence GTIN
      HTS057M 04053252338083