HTS110M

ChemiSCREEN Membrane Preparation Recombinant Human 5-HT_4B Serotonin Receptor

Human 5-HT4 GPCR membrane preparation for Radioligand binding Assays & GTPγS 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_000870
• Numéro d'accès UniProt: Q8IXH9
• Conditions d'expédition: dry ice

Description

Description générale

5-Hydroxytryptamine (5-HT, also commonly known as serotonin) is synthesized in enterochromaffin cells in the intestine and in serotonergic nerve terminals. In the periphery, 5-HT mediates gastrointestinal motility, platelet aggregation, and contraction of blood vessels. Many functions of the central nervous system are influenced by 5-HT, including sleep, motor activity, sensory perception, arousal and appetite. A family of 12 GPCRs and one ion channel mediate the biological effects of 5-HT (Hoyer et al., 1994). 5-HT₄ comprises at least 8 isoforms varying at the C-terminus, which are generated by alternative splicing. The expression and distribution of these splice variants differs among organs and tissues with many of them present in several tissues such as atrium, brain, and GI tract (Bockaert et al. 2004). To date, all isoforms have been shown to activate adenylyl cyclase in vitro, and no difference in signal transduction between C-terminal 5-HT₄ receptor variants has been demonstrated. 5-HT₄ receptors are of potential interest for the treatment of patients with GI motility disorders and Alzheimer′s disease (De Maeyer et al. 2008; Lezoualc’h 2007). Millipore′s 5-HT_4B 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 agonists and antagonists of 5-HT_4B. The membrane preparations exhibit a Kd of 0.46 nM for [³H]-GR113808. With 1 nM [³H]-GR113808, 5 μg/well 5-HT4B Membrane Prep yields greater than 10 fold signal-to-background ratio.

Human HTR4, transcript b, encoding 5-HT_4B

Application

Human 5-HT4 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Radioligand binding assay and GTPγS binding.

Actions biochimiques/physiologiques

GPCR Class: A

Protein Target: 5-HT4

Target Sub-Family: Serotonin

Qualité

Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT_4B membrane prep.

	10 µg/well	5 µg/well
Signal:Background	26.4	21.7
Specific Binding (cpm)	5286	4879

SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 12.5 pmol/mg
K_d: 2.7 nM for [³H]-GR113808

Caractéristiques

Inucbation Conditions
RECOMMENDED ASSAY CONDITIONS: Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in assay 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 assay buffer. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with assay buffer. The plate is dried and counted.

Assay buffer: 25 mM Tris, pH 7.4, filtered and stored at 4°C

Radioligand: [³H]-GR113808 (Amersham TRK944)

One package contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 10-fold signal:background with ³H-labeled GR113808 at 1 nM.

Forme physique

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.

Stockage et stabilité

Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.

Informations légales

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Clause de non-responsabilité

Ce produit, destiné à la recherche scientifique, est soumis à une réglementation spécifique en France, y compris pour les activités d′importation et d′exportation (Article L 1211-1 alinéa 2 du Code de la Santé Publique). L′acheteur (c′est-à-dire l′utilisateur FINAL) est tenu d′obtenir une autorisation d′importation auprès du ministère français de la recherche, mentionné à l′article L1245-5-1 II du Code de la Santé Publique. En commandant ce produit, vous confirmez détenir l′autorisation d′importation requise.

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