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HTS181M ChemiSCREEN™ Membrane Preparation Recombinant Human NPBW2 Neuropeptide B/W Receptor

Human NPBW2 / GPR8 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: P48146 Entrez Gene Number: NM_005286.2
HTS181M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1NPBW2 / GPR8
Description
Catalogue NumberHTS181M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NPBW2 Neuropeptide B/W Receptor
OverviewFull length human GPR8 cDNA encoding NPBW2
Background InformationNeuropeptide B (NPB) and neuropeptide W (NPW) are members of a recently identified neuropeptide family that are ligands for NPBW1 and NPBW2 receptors, both of which are coupled to Gi/o protein to inhibit intracellular cAMP production and share 64% sequence homology (Singh and Davenport, 2006). NPBW1 recognizes both NPB and NPW with similar nanomolar affinities (with a slight preference for NPB), whereas NPBW2 is moderately selective for NPW (Tanaka et al., 2003). NPBW2 is one of a few GPCRs that have no rat or mouse orthologue, although gene encoding NPBW2 has been discovered in other mammalian species such as rabbit (Lee et al., 1999). NPBW2 mRNA is known to be expressed in the frontal cortex, parietal cortex hippocampus, caudate nucleus, thalamus, pituitary, adrenal gland, and lymph node (Brezillon et al., 2003). Functions of NPBW2 may be involved in feeding, weight regulation, and pain response through direct or indirect actions in the central nervous system. Millipore's NPBW2 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 NPBW2 receptor interactions with its ligand. The membrane preparations exhibit a Kd of 2nM for [125I]-Neuropeptide W23. With 10 ug/well NPBW2 Membrane Prep and 0.35nM [125I]-Neuropeptide W23, a greater than 10-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 was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Quality LevelMQ100
Applications
ApplicationHuman NPBW2 / GPR8 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 SummaryThe protein encoded by this gene is an integral membrane protein and G protein-coupled receptor. The encoded protein is similar in sequence to another G protein-coupled receptor (GPR7), and it is structurally similar to opioid and somatostatin receptors. This protein binds neuropeptides B and W. This gene is intronless and is expressed primarily in the frontal cortex of the brain.
Gene Symbol
  • NPBWR2
  • GPR8
GPCR ClassA
Host CellsChem-1
Protein TargetNPBW2 / GPR8
Target Sub-FamilyNeuropeptide B/W
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P48146 # Interacts specifically with a number of opioid ligands. Receptor for neuropeptides B and W, which may be involved in neuroendocrine system regulation, food intake and the organization of other signals.
SIZE: 333 amino acids; 36861 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Detected at high levels in caudate nucleus, hyppocampus and amygdala; at moderate levels in the adult brain, thalamus, parietal cortex, pituitary gland, adrenal gland and lymph nodes.
SIMILARITY: SwissProt: P48146 ## 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 Tris, pH 7.4. 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, filtered and stored at 4°C.
Radioligand: [125I]-neuropeptide W23 (Perkin Elmer#: NET-403)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 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 10-fold signal:background with 125I-labeled neuropeptide W23 at 0.35nM.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with 10 or 20 µg/well of NPBW2 Receptor membrane prep.
Product Usage Statements
20 µg/well 10 µg/well
Signal:Background 26.3 30.2
Specific Binding (cpm) 20148 19282


SPECIFICATIONS: 1 unit = 10.0 µg membrane preparation
Bmax: 11.34 pmol/mg
Kd: 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
Global Trade Item Number
Catalogue Number GTIN
HTS181M 04053252672699

Documentation

References

Reference overviewPub Med ID
Neuropeptide B and W: neurotransmitters in an emerging G-protein-coupled receptor system.
Singh, Gurminder and Davenport, Anthony P
Br. J. Pharmacol., 148: 1033-41 (2006)  2006

Show Abstract 		16847439 16847439
Identification of natural ligands for the orphan G protein-coupled receptors GPR7 and GPR8
Brezillon, Stephane, et al
J Biol Chem, 278:776-83 (2003)  2003

12401809 12401809
Characterization of a family of endogenous neuropeptide ligands for the G protein-coupled receptors GPR7 and GPR8
Tanaka, Hirokazu, et al
Proc Natl Acad Sci USA, 100:6251-6 (2003)  2003

12719537 12719537

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