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HTS079M ChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation

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HTS079M
200 units  
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      Overview

      Replacement Information

      Key Spec Table

      SpeciesHost CellsProtein Target
      Human Chem-1VPAC2 / VIP2
      Description
      Catalogue NumberHTS079M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation
      OverviewHuman VPAC2
      Background InformationVasoactive intestinal peptide (VIP), a 28 amino acid peptide originally isolated by its vasodilation activity, binds to two class B GPCRs, VPAC1 and VPAC2, to exert its functions in the CNS, vasculature, immune system and adrenal medulla (Harmar et al., 1998). In the immune system, VPAC2 is expressed on stimulated CD4 T cells, and binding of T cell-derived VIP to VPAC2 induces a shift toward the Th2 pathway (Delgado et al., 2004; Voice et al., 2004). In addition, VPAC2 is an essential regulator of the circadian pacemaker of the hypothalamic suprachiasmatic nuclei (Hughes et al., 2004). Chemicon's VPAC2 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 VPAC2 interactions with PACAP. The membrane preparations exhibit a Kd of 0.9 nM for [125I]-PACAP27. With 5 µg/well VPAC2 Membrane Prep and 0.5 nM [125I]-PACAP27, a greater than 15-fold signal-to-background ratio was obtained.
      References
      Product Information
      FormatMembranes
      PresentationLiquid 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 VPAC2 / VIP2 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 SummaryVasoactive intestinal peptide (VIP; MIM 192320) and pituitary adenylate cyclase activating polypeptide (PACAP; MIM 102980) are homologous peptides that function as neurotransmitters and neuroendocrine hormones. While the receptors for VIP and PACAP share homology, they differ in their substrate specificities and expression patterns. See VIPR1 (MIM 192321) and ADCYAP1R1(MIM 102981).[supplied by OMIM]
      Gene Symbol
      • VIPR2
      • VPCAP2R
      • VPAC2
      • VIP2R
      • PACAP-R-3
      • FLJ16511
      • VIP-R-2
      GPCR ClassB
      Host CellsChem-1
      Protein TargetVPAC2 / VIP2
      Target Sub-FamilyVIP/PACAP
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P41587 # This is a receptor for VIP as well as PACAP-38 and -27, the activity of this receptor is mediated by G proteins which activate adenylyl cyclase. Can be coupled to phospholipase C.
      SIZE: 438 amino acids; 49479 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P41587 ## Belongs to the G-protein coupled receptor 2 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] PACAP27 (Perkin Elmer # NEX294)
      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 an unit is the amount of membrane that will yield greater than 15-fold signal:background with 125I-labeled PACAP27 at 0.5 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 5ug/well of VPAC2 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 25.5. 22.8
      Specific Binding (cpm) 32913 27650


      SPECIFICATIONS: 1 unit = 5 µg membrane preparation
      Bmax: 17.3 pmol/mg
      Kd: 0.9 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
      Catalogue Number GTIN
      HTS079M 04053252406928

      Documentation

      ChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      ChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation Certificates of Analysis

      TitleLot Number
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN VPAC2 VIP/PACAP RECEPTOR - 2326017 2326017

      References

      Reference overviewPub Med ID
      Aberrant gating of photic input to the suprachiasmatic circadian pacemaker of mice lacking the VPAC2 receptor
      Hughes, Alun T, et al
      J Neurosci, 24:3522-6 (2004)  2004

      15071099 15071099
      The significance of vasoactive intestinal peptide in immunomodulation
      Delgado, Mario, et al
      Pharmacol Rev, 56:249-90 (2004)  2004

      15169929 15169929
      c-Maf and JunB mediation of Th2 differentiation induced by the type 2 G Protein-Coupled Receptor (VPAC2) for vasoactive intestinal peptide.
      Voice J et al.
      J. Immunol., 172:7289-7296 (2004)  2004

      15187104 15187104
      International Union of Pharmacology. XVIII. Nomenclature of receptors for vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide
      Harmar, A J, et al
      Pharmacol Rev, 50:265-70 (1998)  1998

      9647867 9647867

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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