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HTS066M ChemiSCREEN™ Human Y2 Neuropeptide Receptor Membrane Preparation

HTS066M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1Y2
Description
Catalogue NumberHTS066M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Human Y2 Neuropeptide Receptor Membrane Preparation
OverviewFull-length human NPY2R cDNA encoding Y2
Background InformationThe NPY family consists of three 36-amino acid peptides, neuropeptide Y (NPY), peptide YY (PYY) and pancreatic polypeptide (PP), which bind to the NPY receptor family of G protein-coupled receptors. Five NPY receptors, Y1, Y2, Y4, Y5 and Y6, have been defined at the molecular level, and each signals primarily through Gi/o. Binding of NPY family peptides to NPY receptors mediates a variety of physiological effects, including promotion of food intake, decreased anxiety, inhibition of neurotransmitter and hormone release, vasoconstriction, and gut motility. Y2 is primarily expressed in the CNS, and it mediates presynaptic inhibition of neurotransmitter release (Michel et al., 1998). Chemicon's Y2 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 Y2 interactions and its ligands. The membrane preparations exhibit a Kd of 1.28 nM for [125I]-human PYY. With 0.5 nM [125I]-human PYY, 5 or 10µg/well Y2 Membrane Prep yields greater than 10-fold signal-to-background ratio.
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 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman Y2 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
Gene Symbol
  • NPY2R
  • NPY2-R
GPCR ClassA
Host CellsChem-1
Protein TargetY2
Target Sub-FamilyNeuropeptide Y
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P49146 # Receptor for neuropeptide Y and peptide YY. The rank order of affinity of this receptor for pancreatic polypeptides is PYY > NPY > PYY (3-36) > NPY (2-36) > [Ile-31, Gln-34] PP > [Leu- 31, Pro-34] NPY > PP, [Pro-34] PYY and NPY free acid.
SIZE: 381 amino acids; 42731 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: High levels in amygdala, corpus callosum, hippocampus and subthalamic nucleus. Also detectable in caudate nucleus, hypothalamus and substantia nigra.
SIMILARITY: SwissProt: P49146 ## 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, 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]-human PYY (Perkin Elmer#:NEX-341 )
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 10-fold signal:background with 125I labeled human PYY 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 varying amounts of TP membrane prep:
Product Usage Statements
10 µg/well 5 µg/well
Signal:Background 19.8 26.3
Specific Binding (cpm) 27347 23335


SPECIFICATIONS: 1 unit = 5 µg
Bmax for [125I] human PYY binding: 17.89 pmol/mg protein;
Kd for [125I] human PYY binding: ~ 1.28 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
HTS066M 04053252510090

Documentation

ChemiSCREEN™ Human Y2 Neuropeptide Receptor Membrane Preparation SDS

Title

Safety Data Sheet (SDS) 

References

Reference overviewPub Med ID
XVI. International Union of Pharmacology recommendations for the nomenclature of neuropeptide Y, peptide YY, and pancreatic polypeptide receptors.
Michel, M C, et al.
Pharmacol. Rev., 50: 143-50 (1998)  1998

9549761 9549761

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Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations