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ChemiScreen™ Membrane Prep Hu Recombinant VPAC

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货号：HTS043M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 VPAC1 / VIP1
Description
Catalogue Number HTS043M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiScreen™ VPAC1 Membrane Preparation
Overview Human VIPR1 cDNA, encoding VPAC1
Background Information Vasoactive 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
Format Membranes
Presentation Liquid 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
Application Human VPAC1 / VIP1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration 1 mL, 0.5 mg/mL per vial.
Entrez Gene Number
Entrez Gene Summary This 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 Class B
Host Cells Chem-1
Protein Target VPAC1 / VIP1
Target Sub-Family VIP/PACAP
UniProt Number
UniProt Summary FUNCTION: 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 Conditions Membranes 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
1
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of VPAC 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 Conditions Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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