HTS132M

ChemiSCREEN Human 5-HT_2C Receptor Membrane Preparation

Human 5-HT2C GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

• UNSPSCコード: 41106514
• eCl@ss: 32161000
• NACRES: NA.84

特性

• 由来生物: human
• 品質水準: 100
• リコンビナント: expressed in Chem-1 cells
• メーカー／製品名: ChemiScreen; Chemicon^®
• テクニック: ligand binding assay: suitable (GTPγS); radioligand binding assay (RLBA): suitable
• NCBIアクセッション番号: NM_000868.1
• UniProtアクセッション番号: P28335
• 輸送温度: dry ice
• 遺伝子情報: human ... HTR2C(3358)

詳細

詳細

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-HT_2C, which couples to G_q in most cells to stimulate intracellular calcium, is prominently expressed in brain and appears to modulate depression, anxiety and appetite (Miller, 2005; Serretti et al., 2004; Wood, 2003). The mRNA encoding 5-HT_2C undergoes selective RNA editing that changes 4 amino acids in the second intracellular loop; these changes result in alteration of efficiency of coupling to G proteins. Alterations in editing of 5-HT_2C have been detected in victims of suicidal depression and in mice treated with the SSRI, fluoxetine (Tohda et al., 2006). Millipore′′s 5-HT_2C 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 5-HT_2C interactions and its ligands. The membrane preparations exhibit a Kd of 5.9 nM for [³H]-mesulergine. With 6 nM [³H]- Mesulergine, 10 µg/well 5-HT_2C Membrane Prep yields greater than 5 fold signal-to-background ratio.

Human 5-HT_2C

アプリケーション

Radioligand binding assay and GTPγS binding.

生物化学的／生理学的作用

GPCR Class: A

Protein Target: 5-HT2C

Target Sub-Family: Serotonin

品質

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT_2C membrane prep:

	10 µg/well
Signal:Background	7.8
Specific Binding (cpm)	2433

SPECIFICATIONS: 1 unit = 10 µg membrane preparation
Bmax: 15.66 pmol/mg
Kd: 5.9 nM

規格

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, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, 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 Hepes, pH 7.4, 5 mM MgCl₂, 1 mM CaCl₂, filtered and stored at 4°C
Radioligand: [³H]-Mesulergine (Amersham TRK 845)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, filtered and stored at 4°C.

物理的形状

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.

保管および安定性

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.

法的情報

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

免責事項

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.

安全性情報

• 保管分類コード: 12 - Non Combustible Liquids
• WGK: WGK 2
• 引火点(°F): Not applicable
• 引火点(℃): Not applicable