ChemiSCREEN Membrane Preparation Recombinant Human 5-HT₆Serotonin Receptor

半岛bd体育手机客户端 说明

一般描述

Full-length human HTR6 cDNA encoding the 5-HT₆Serotonin Receptor
The neurotransmitter serotonin/5-hydroxytryptamine (5-HT) regulates a wide variety of neurological functions. A family of 13 receptors (12 GPCRs and one ion channel) mediate the effects of serotonin. The serotonin receptor 5-HT₆is a Gs coupled receptor expressed solely in the CNS, primarily in the limbic and cortical regions. 5-HT₆appears to play a role in memory and learning, obesity, psychosis, anxiety and epilepsy (Woolley et al., 2004; Fisas et al., 2006). In particular, a 5-HT₆-selective agonist caused significant weight loss in a rat model of diet-induced obesity. Millipore′s 5-HT₆membrane 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-HT₆. The membrane preparations exhibit a Kd of 0.44 nM for [¹²⁵I]-SB258585. With 0.25nM [¹²⁵I]-SB258585, 5 μg/well 5-HT₆Membrane Prep typically yields greater than 6-fold signal-to-background ratio.

应用

Radioligand binding assay and GTPγS binding.

生化/生理作用

Target Sub-Family: Serotonin
GPCR Class: A
Protein Target: 5-HT6

质量

Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT₆Receptor membrane prep.

	5 ?g/well	10 ?g/well
Signal:Background	12.8	12.3
Specific Binding (cpm)	10192	12675

SPECIFICATIONS: 1 unit = 5 ?g membrane preparation
B _maxfor [ ¹²⁵I]-SB258585 binding: 6.1 pmol/mg protein
K _dfor [ ¹²⁵I]-SB258585 binding: ~0.44 nM

半岛bd体育手机客户端 规格

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 the binding buffer. 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: 20 mM HEPES, 3 mM MgCl₂, 2 mM Ascorbic acid, pH 7.4, filtered and stored at 4°C

Radioligand: [¹²⁵I]-SB258585 (Perkin Elmer #:NEX424)

Wash Buffer: same as the binding buffer.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 6-fold signal:background with¹²⁵I labeled SB258585 at 0.25 nM

外形

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

储存及稳定性

Maintain frozen at –70°C up to expiration date indicated on the label. Do not freeze and thaw.

法律信息

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

免责声明

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.

基本信息

eCl@ss

32161000

半岛bd体育手机客户端 性质

质量水平	100
生物来源	human
重组	expressed in Chem-1 cells
manufacturer/tradename	ChemiScreen Chemicon?
technique(s)	ligand binding assay: suitable (GTPγS) radioligand binding assay (RLBA): suitable
NCBI登记号	NM_000871
UniProt登记号	P50406
运输	dry ice

安全信息

储存分类代码	12 - Non Combustible Liquids
WGK	WGK 2
闪点(F)	Not applicable
闪点(C)	Not applicable