HTS066M
ChemiSCREEN Human Y 2 Neuropeptide Receptor Membrane Preparation
Human Y2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Code UNSPSC :
41106514
eCl@ss :
32161000
Nomenclature NACRES :
NA.84
Produits recommandés
Source biologique
human
Niveau de qualité
Produit recombinant
expressed in Chem-1 cells
Fabricant/nom de marque
ChemiScreen
Chemicon®
Technique(s)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
Numéro d'accès NCBI
Numéro d'accès UniProt
Conditions d'expédition
dry ice
Informations sur le gène
human ... NPY2R(4887)
Description générale
Full-length human NPY2R cDNA encoding Y2
The 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.
Application
Human Y2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding.
Actions biochimiques/physiologiques
GPCR Class: A
Protein Target: Y2
Target Sub-Family: Neuropeptide Y
Qualité
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of TP membrane prep:
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
| 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
Caractéristiques
Inucbation 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]-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
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]-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
Forme physique
Liquid 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.
Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 mL packaging buffer, rapidly frozen, and stored at -80°C.
Stockage et stabilité
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Informations légales
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Clause de non-responsabilité
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.
Code de la classe de stockage
10 - Combustible liquids
Classe de danger pour l'eau (WGK)
WGK 2
Certificats d'analyse (COA)
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
XVI. International Union of Pharmacology recommendations for the nomenclature of neuropeptide Y, peptide YY, and pancreatic polypeptide receptors.
M C Michel et al.
Pharmacological reviews, 50(1), 143-150 (1998-04-29)
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