Millipore Sigma Vibrant Logo
Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

HTS079M ChemiSCREEN™ Human VPAC2 VIP Receptor Membrane Preparation

Human VPAC2 / VIP2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P41587 Entrez Gene Number: NM_003382.3
HTS079M
200 units  
Purchase on Sigma-Aldrich

Overview

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.
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.
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
Global Trade Item Number
Catalogue Number GTIN
HTS079M 04053252406928