HTS182M ChemiSCREEN™ Membrane Preparation Recombinant Human PK2 Prokineticin Receptor

HTS182M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1 PK2 / PKR2
      Description
      Catalogue NumberHTS182M
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human PK2 Prokineticin Receptor
      OverviewFull-length human GPR73L1 cDNA encoding PK2
      Background InformationProkineticins, also known as endocrine gland vascular endothelial growth factors (EG-VEGF), are two ~10 kD secreted proteins originally described to mediate angiogenesis and gastrointestinal smooth muscle contraction (Li et al., 2001; LeCouter et al., 2003). Subsequently, prokineticins have been found to mediate central nervous system functions including circadian rhythms and olfactory bulb development (Cheng et al., 2002; Ng et al., 2005). Two Gq-coupled receptors, PK1 and PK2 (also known as GPR73a and GPR73b), mediate cellular responses to prokineticins (Lin et al., 2002). Millipore's PK2 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 agonists and antagonists at PK2. The membrane preparations exhibit a Kd of 0.36 nM for [125I]-MIT-1. With 0.3 nM [125I]-MIT-1, 10µg/well PK2 Membrane Prep typically yields greater than 5-fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationOne package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 5-fold signal:background with 125I-labeled MIT-1 at 0.3 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 1 mg/mL in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C.
      Applications
      ApplicationHuman PK2 / PKR2 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 SummaryProkineticins are secreted proteins that can promote angiogenesis and induce strong gastrointestinal smooth muscle contraction. The protein encoded by this gene is an integral membrane protein and G protein-coupled receptor for prokineticins. The encoded protein is similar in sequence to GPR73, another G protein-coupled receptor for prokineticins.
      Gene Symbol
      • PROKR2
      • PKR2
      • GPR73L1
      • GPR73b
      • PK-R2
      • dJ680N4.3
      • OTTHUMP00000030192
      • KAL3
      • GPRg2
      GPCR ClassA
      Host CellsChem-1
      Protein TargetPK2 / PKR2
      Target Sub-FamilyProkineticin
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: Q8NFJ6 # Receptor for prokineticin 2. Exclusively coupled to the G(q) subclass of heteromeric G proteins. Activation leads to mobilization of calcium, stimulation of phosphoinositide turnover and activation of p44/p42 mitogen-activated protein kinase.
      SIZE: 384 amino acids; 43996 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: Expressed in the ileocecum, thyroid gland, pituitary gland, salivary gland, adrenal gland, testis, ovary and brain.
      DISEASE: SwissProt: Q8NFJ6 # Defects in PROKR2 are the cause of Kallmann syndrome type 3 (KAL3) [MIM:244200]; also known as hypogonadotropic hypogonadism and anosmia. Anosmia or hyposmia is related to the absence or hypoplasia of the olfactory bulbs and tracts. Hypogonadism is due to deficiency in gonadotropin-releasing hormone and probably results from a failure of embryonic migration of gonadotropin- releasing hormone-synthesizing neurons. KAL3 patients have variable degrees of olfactory and reproductive dysfunction, but do not show any of the occasional clinical anomalies reported in Kallmann syndrome such as renal agenesis, cleft lip and/or palate, selective tooth agenesis, and bimanual synkinesis.
      SIMILARITY: SwissProt: Q8NFJ6 ## 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, 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: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
      Radioligand: [125I] MIT-1 (Perkin Elmer#: NEX-410)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      10 µg/well
      Signal:Background 9.80
      Specific Binding (cpm) 6874
      SPECIFICATIONS: 1 unit = 10 µg
      Bmax for [125I]- MIT-1 binding: 0.52 pmol/mg protein
      Kd for [125I]- MIT-1 binding: ~0.36 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

      Documentation

      ChemiSCREEN™ Membrane Preparation Recombinant Human PK2 Prokineticin Receptor SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Dependence of olfactory bulb neurogenesis on prokineticin 2 signaling
      Ng, Kwan L, et al
      Science, 308:1923-7 (2005)  2005

      15976302 15976302
      Prokineticin 2 transmits the behavioural circadian rhythm of the suprachiasmatic nucleus.
      Cheng, Michelle Y, et al.
      Nature, 417: 405-10 (2002)  2002

      Show Abstract
      12024206 12024206
      Endocrine gland-derived VEGF and the emerging hypothesis of organ-specific regulation of angiogenesis.
      LeCouter, Jennifer, et al.
      Nat. Med., 8: 913-7 (2002)  2002

      Show Abstract
      12205443 12205443

      Data Sheet

      Title
      ChemiSCREEN Membrane Preparation Recombinant Human PK2 Prokineticin Receptor - Data Sheet

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      Categories

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