HTS108M

ChemiSCREEN Membrane Preparation Recombinant Human 5-HT_1B Serotonin Receptor

Human 5-HT1B GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

• UNSPSC Code: 41106514
• eCl@ss: 32161000

Propiedades

• biological source: human
• Quality Level: 100
• recombinant: expressed in Chem-1 cells
• manufacturer/tradename: ChemiScreen; Chemicon^®
• technique(s): ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• NCBI accession no.: NM_000863.1
• UniProt accession no.: P28222
• shipped in: dry ice

Descripción

General description

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). The 5-HT_1B receptor (also known as 5-HT_1Dβ) is expressed presynaptically on serotonergic neurons and postsynaptically on non-serotonergic neurons, and it regulates release of 5-HT and acetylcholine. Several brain regions, including globus pallidus, substantia nigra and dorsal subiculum, highly express HT1B. 5-HT_1B has been implicated in the physiology of aggression, depression, migraine, anxiety and reward for drugs of abuse (Sari, 2004). 5-HT_1B interacts with p11, which increases cell surface expression and function of 5-HT_1B (Svenningsson et al., 2006). Millipore′s 5-HT_1B 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_1B. The membrane preparations exhibit a Kd of 10.5 nM for [³H]-GR125743. With 7 nM [³H]-GR125743, 10μg/well 5-HT_1B Membrane Prep typically yields greater than 8-fold signal-to-background ratio.

Full-length human HTR1B cDNA encoding the 5-HT_1B Serotonin Receptor

Application

Radioligand binding assay and GTPγS binding.

Biochem/physiol Actions

GPCR Class: A

Protein Target: 5-HT1B

Target Sub-Family: Serotonin

Quality

Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst5 Receptor membrane prep.

	10 µg/well	20 µg/well
Signal:Background	12.4	10.5
Specific Binding (cpm)	3210	3352

SPECIFICATIONS: 1 unit =10 μg
Bmax for [³H]-GR125743 binding: 3.7 pmol/mg protein
Kd for [³H]-GR125743 binding: ~10.5 nM

Specifications

Inucbation Conditions
RECOMMENDED ASSAY 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, 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C

Radioligand: [³H]-GR125743. (Amersham #:TRK-1046 )

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 a unit is the amount of membrane that will yield greater than 8-fold signal:background with ³H labeled GR125743 at 7 nM

Physical form

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
Packaging method: Membranes protein were adjusted to 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.

Storage and Stability

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Legal Information

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

Disclaimer

RESEARCH USE ONLY. This product is regulated in France when intended to be used for scientific purposes, including for import and export activities (Article L 1211-1 paragraph 2 of the Public Health Code). The purchaser (i.e. enduser) is required to obtain an import authorization from the France Ministry of Research referred in the Article L1245-5-1 II. of Public Health Code. By ordering this product, you are confirming that you have obtained the proper import authorization.

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.

Información de seguridad

• Storage Class: 12 - Non Combustible Liquids
• wgk_germany: WGK 2
• flash_point_f: Not applicable
• flash_point_c: Not applicable