ChemiSCREEN Membrane Preparation Recombinant Human 5-HT₇Serotonin Receptor

半岛bd体育手机客户端 说明

一般描述

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-HT? is a Gs coupled receptor expressed in the brain (hypothalamus, hippocampus and frontal cortex) and in smooth muscle and GI tract. Systemic delivery of serotonin results in hypothermia, and pharmacological and genetic studies indicate that 5-HT? mediates this effect (Hedlund et al., 2003). In addition, 5-HT? appears to modulate circadian rhythms and smooth muscle relaxation, and is implicated as a target for sleep, particularly as related to depression, and migraine (Terron and Falcon-Neri, 1999; Mullins et al., 1999). Millipore′s 5-HT? 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?. The membrane preparations exhibit a Kd of 37.7 nM for [?H]-5-Carboxamidotryptamine (5-CT). With 30 nM [?H]-5-CT, 10 μg/well 5-HT? Membrane Prep typically yields greater than 4-fold signal-to-background ratio.
Full-length human HTR7 cDNA encoding the 5-HT? Serotonin Receptor

应用

Human 5-HT7 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

生化/生理作用

Target Sub-Family: Serotonin
GPCR Class: A
Protein Target: 5-HT7

质量

Table 1. Signal:background and specific binding values obtained in a competition binding assay with 10 μg of 5-HT? Receptor membrane prep.

	10 ?g/well
Signal:Background	7.5
Specific Binding (cpm)	545

SPECIFICATIONS: 1 unit =10 μg
Bmax for [?H]-5-CT binding: 20.1 pmol/mg protein
Kd for [?H]-5-CT binding: ~37.7 nM

半岛bd体育手机客户端 规格

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, 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: 50 mM Tris, pH 7.4, 5 mM MgCl?, 4 mM CaCl?, 0.5 mM ascorbic acid, filtered and stored at 4°C

Radioligand: [?H]-5-Carboxamidotryptamine (Perkin Elmer NET1071)

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 4-fold signal:background with ?H-labeled 5-CT at 30 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 for up to 2 years. 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_000872
UniProt登记号	P34969
运输	dry ice
Gene Information	human ... HTR7(3363)

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