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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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HTS132M
Sigma-AldrichChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation
Human 5-HT2C GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
ChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation
Overview
Human 5-HT2C
Background Information
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.
References
Product Information
Format
Membranes
HS Code
3822 19 90
Presentation
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.
Serotonin (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.
FUNCTION: 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 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Signal:background and specific binding values obtained in a competition binding assay with varying amounts of 5-HT2C membrane prep:
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 Conditions
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.
Packaging Information
Material Size
200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
HTS132M
04053252624759
Documentation
ChemiSCREEN™ Human 5-HT2C Receptor Membrane Preparation SDB
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
Anxiety disorders are the most common psychiatric illness affecting both adults and children. Following the observation that m-chlorophenylpiperazine(mCPP) induced anxiety-like states in patients and in animal models, it was shown that in man, mCPP behaves as a functionally selective agonist at the 5-hydroxytryptamine (5-HT)2C receptor. This caused much interest in the development of antagonists at the 5-HT2C receptor for the treatment of anxiety disorders. This review examines the pre-clinical and clinical evidence for a role of the 5-HT2C receptor in anxiety and evaluates the progress of compounds that target this therapeutic approach.
Millipore offers a large selection of robust and reliable G-protein coupled receptor products, including Stable Cell Lines, Membrane Preparations, and Frozen Cells. See below for GPCR research tools. Weitere Informationen >>
