HTS102M

ChemiScreen D₁ Dopamine Receptor Membrane Preparation

Human D1 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

• Code UNSPSC : 41106514
• eCl@ss : 32161000
• Nomenclature NACRES : NA.84

Propriétés

• Source biologique: human
• Niveau de qualité: 100
• 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: NM_000794.3
• Numéro d'accès UniProt: P21728
• Conditions d'expédition: dry ice
• Informations sur le gène: human ... DRD1(1812)

Description

Description générale

Dopamine exerts its action by binding to five distinct dopamine receptors, all of which belong to G protein-coupled receptor family (Missale et al. 1998). The D₁ subtype is the most abundant dopamine receptor in the central nervous system. Activation of D₁ receptor stimulates adenylyl cyclase, activates cyclic AMP-dependent protein kinases. It regulates neuronal growth and development, mediate some behavioral responses and modulate dopamine receptor D₂-mediated events. Chemicon′s D₁ 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 H₁ interactions and its ligands. The membrane preparations exhibit a Kd of 1.5 nM for [^{3H]-SCH 23390. With 1.5 nM [3H]- SCH 23390, 2.5 µ}g/well and 10 µg/well D₁ Membrane Prep typically yield greater than 5 and 15-fold signal-to-background ratios, respectively.

Full-length human DRD1 cDNA

Application

Radioligand binding assay and GTPS binding.

Actions biochimiques/physiologiques

GPCR Class: A

Protein Target: D1

Target Sub-Family: Dopamine

Qualité

Bmax: 11.85 pmol/mg; Kd: 1.5 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 Tris, pH 7.4, 10 mM MgCl₂, 1 mM EDTA, filtered and stored at 4°C
Radioligand: [³H] SCH 23390 (Perkin Elmer#:NET-930 )
Wash Buffer: 50 mM Tris, pH 7.4 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 ³H-labeled SCH 23390 at 1.5 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.

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.

Informations sur la sécurité

• Code de la classe de stockage: 12 - Non Combustible Liquids
• Classe de danger pour l'eau (WGK): WGK 2
• Point d'éclair (°F): Not applicable
• Point d'éclair (°C): Not applicable