HTS132M
ChemiSCREEN Human 5-HT2C Receptor Membrane Preparation
Human 5-HT2C GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Code UNSPSC :
41106514
eCl@ss :
32161000
Nomenclature NACRES :
NA.84
Source biologique
human
Niveau de qualité
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
Numéro d'accès UniProt
Conditions d'expédition
dry ice
Informations sur le gène
human ... HTR2C(3358)
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-HT2C, which couples to Gq 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-HT2C 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-HT2C have been detected in victims of suicidal depression and in mice treated with the SSRI, fluoxetine (Tohda et al., 2006). Millipore′′s 5-HT2C 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-HT2C 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-HT2C Membrane Prep yields greater than 5 fold signal-to-background ratio.
Human 5-HT2C
Application
Radioligand binding assay and GTPγS binding.
Actions biochimiques/physiologiques
GPCR Class: A
Protein Target: 5-HT2C
Target Sub-Family: Serotonin
Qualité
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT2C membrane prep:
SPECIFICATIONS: 1 unit = 10 µg membrane preparation
Bmax: 15.66 pmol/mg
Kd: 5.9 nM
| 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
Caractéristiques
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.
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.
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.
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
Certificats d'analyse (COA)
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Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.
Serotonin 5-ht2c receptor agonists: potential for the treatment of obesity
Miller, Keith J
Molecular Interventions, 5, 282-291 (2005)
The 5-HT2C receptor as a target for mood disorders
Serretti, Alessandro, et al
Expert Opinion on Therapeutic Targets, 8, 15-23 (2004)
N M Barnes et al.
Neuropharmacology, 38(8), 1083-1152 (1999-08-26)
It is now nearly 5 years since the last of the currently recognised 5-HT receptors was identified in terms of its cDNA sequence. Over this period, much effort has been directed towards understanding the function attributable to individual 5-HT receptors
Therapeutic potential of 5-HT2C receptor antagonists in the treatment of anxiety disorders.
Wood, Martyn D
Current Drug Targets. CNS and Neurological Disorders, 2, 383-387 (2003)
Molecular pathopharmacology of 5-HT2C receptors and the RNA editing in the brain
Tohda, Michihisa, et al
Journal of Pharmacological Sciences, 100, 427-432 (2006)
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