HTS043M ChemiScreen™ VPAC1 Membrane Preparation

HTS043M
200 units  
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      Overview

      Replacement Information

      Key Specifications Table

      SpeciesHost CellsProtein Target
      Human Chem-1 VPAC1 / VIP1
      Description
      Catalogue NumberHTS043M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiScreen™ VPAC1 Membrane Preparation
      OverviewHuman VIPR1 cDNA, encoding VPAC1
      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, VIP is synthesized by mast cells and lymphocytes, and appears to inhibit inflammation and to shift the immune response toward a Th2 pathway (Delgado et al., 2004). In the heart, VIP is expressed by nerve fibers, where it modulates heart rate, and coronary blood flow (Henning and Sawmiller, 2001). Chemicon's VPAC1 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 VPAC1 interactions with VIP. The membrane preparations exhibit a Kd of 1.2 nM for [125I]-VIP. With 2.5 μg/well VPAC1 Membrane Prep and 0.4 nM [125I]-VIP, a greater than 5-fold signal-to-background ratio is obtained.
      References
      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 the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
      Applications
      ApplicationHuman VPAC1 / VIP1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay, and GTPgammaS binding.
      Biological Information
      Concentration1 mL, 0.5 mg/mL per vial.
      Entrez Gene Number
      Entrez Gene SummaryThis gene encodes a receptor for vasoactive intestinal peptide, a small neuropeptide. Vasoactive intestinal peptide is involved in smooth muscle relaxation, exocrine and endocrine secretion, and water and ion flux in lung and intestinal epithelia. Its actions are effected through integral membrane receptors associated with a guanine nucleotide binding protein which activates adenylate cyclase.
      Gene Symbol
      • VIPR1
      • HVR1
      • PACAP-R-2
      • RCD1
      • VPCAP1R
      • II
      • FLJ41949
      • VIP-R-1
      • VAPC1
      • VPAC1
      • VIPR
      • VIRG
      • RDC1
      GPCR ClassB
      Host CellsChem-1
      Protein TargetVPAC1 / VIP1
      Target Sub-FamilyVIP/PACAP
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P32241 # This is a receptor for VIP. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase. The affinity is VIP = PACAP-27 > PACAP-38.
      SIZE: 457 amino acids; 51547 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      TISSUE SPECIFICITY: In lung, HT29 colonic epithelial cells, Raji B-lymphoblasts. Lesser extent in brain, heart, kidney, liver and placenta.
      SIMILARITY: SwissProt: P32241 ## 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] VIP (Perkin Elmer# NEX192)

      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 5-fold signal:background with 125I-labeled VIP at 0.4 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 varying amounts of VPAC1 membrane prep:
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 9.7 10.8
      Specific Binding (cpm) 30299 31224


      SPECIFICATIONS:
      1 unit = 5 µg
      Bmax: 19.65 pmol/mg
      Kd: 1.2 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™ VPAC1 Membrane Preparation SDS

      Title

      Safety Data Sheet (SDS) 

      References

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

      15169929 15169929
      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

      Data Sheet

      Title
      ChemiScreen VPAC1 Membrane Preparation - 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