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HTS132M ChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation

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

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P28335 Entrez Gene Number: NM_000868.1
HTS132M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-15-HT2C
Description
Catalogue NumberHTS132M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation
OverviewHuman 5-HT2C
Background Information5-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.
References
Product Information
FormatMembranes
PresentationLiquid 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.
Quality LevelMQ100
Applications
ApplicationHuman 5-HT2C GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPγS binding.
Biological Information
Entrez Gene Number
Entrez Gene SummarySerotonin (5-hydroxytryptamine, 5-HT), a neurotransmitter, elicits a wide array of physiological effects by binding to several receptor subtypes, including the 5-HT2 family of seven-transmembrane-spanning, G-protein-coupled receptors, which activate phospholipase C and D signaling pathways. This gene encodes the 2C subtype of serotonin receptor and its mRNA is subject to multiple RNA editing events, where genomically encoded adenosine residues are converted to inosines. RNA editing is predicted to alter amino acids within the second intracellular loop of the 5-HT2C receptor and generate receptor isoforms that differ in their ability to interact with G proteins and the activation of phospholipase C and D signaling cascades, thus modulating serotonergic neurotransmission in the central nervous system. Studies in humans have reported abnormalities in patterns of 5-HT2C editing in depressed suicide victims.
Gene Symbol
  • HTR2C
  • 5-HT-2C
  • 5-HT2C
  • HTR1C
  • 5HT-1C
  • 5-HTR2C
GPCR ClassA
Host CellsChem-1
Protein Target5-HT2C
Target Sub-FamilySerotonin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P28335 # This is one of the several different receptors for 5- hydroxytryptamine (serotonin), a biogenic hormone that functions as a neurotransmitter, a hormone, and a mitogen. This receptor mediates its action by association with G proteins that activate a phosphatidylinositol-calcium second messenger system.
SIZE: 458 amino acids; 51821 Da
SUBUNIT: Interacts with MPDZ.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
DOMAIN: SwissProt: P28335 The PDZ domain-binding motif is involved in the interaction with MPDZ.
PTM: N-glycosylated.
SIMILARITY: Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsMembranes 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT2C membrane prep:
Product Usage Statements
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 Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsStore 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.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS132M 04053252624759

Documentation

ChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation SDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT2C SEROTONIN RECEPTOR - 2010894 2010894
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT2C SEROTONIN RECEPTOR - 2055916 2055916

References

Reference overviewPub Med ID
Molecular pathopharmacology of 5-HT2C receptors and the RNA editing in the brain
Tohda, Michihisa, et al
J Pharmacol Sci, 100:427-32 (2006)  2006

16799258 16799258
Serotonin 5-ht2c receptor agonists: potential for the treatment of obesity
Miller, Keith J
Mol Interv, 5:282-91 (2005)  2005

16249524 16249524
The 5-HT2C receptor as a target for mood disorders
Serretti, Alessandro, et al
Expert Opin Ther Targets, 8:15-23 (2004)  2004

14996615 14996615
Therapeutic potential of 5-HT2C receptor antagonists in the treatment of anxiety disorders.
Wood, Martyn D
Current drug targets. CNS and neurological disorders, 2: 383-7 (2003)  2003

Show Abstract 		14683466 14683466
A review of central 5-HT receptors and their function
Barnes, N M and Sharp, T
Neuropharmacology, 38:1083-152 (1999)  1999

10462127 10462127

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