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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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HTS108M
Sigma-AldrichChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor
Human 5-HT1B GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor
Overview
Full-length human HTR1B cDNA encoding the 5-HT1B Serotonin Receptor
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). The 5-HT1B receptor (also known as 5-HT1Dβ) is expressed presynaptically on serotonergic neurons and postsynaptically on non-serotonergic neurons, and it regulates release of 5-HT and acetylcholine. Several brain regions, including globus pallidus, substantia nigra and dorsal subiculum, highly express HT1B. 5-HT1B has been implicated in the physiology of aggression, depression, migraine, anxiety and reward for drugs of abuse (Sari, 2004). 5-HT1B interacts with p11, which increases cell surface expression and function of 5-HT1B (Svenningsson et al., 2006). Millipore's 5-HT1B 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-HT1B. The membrane preparations exhibit a Kd of 10.5 nM for [3H]-GR125743. With 7 nM [3H]-GR125743, 10μg/well 5-HT1B Membrane Prep typically yields greater than 8-fold signal-to-background ratio.
References
Product Information
Format
Membranes
Presentation
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives. Packaging method: Membranes protein were adjusted to 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
The neurotransmitter serotonin (5-hydroxytryptamine; 5-HT) exerts a wide variety of physiologic functions through a multiplicity of receptors and may be involved in human neuropsychiatric disorders such as anxiety, depression, or migraine. These receptors consist of 4 main groups, 5-HT-1, 5-HT-2, 5-HT-3, and 5-HT4, subdivided into several distinct subtypes on the basis of their pharmacologic characteristics, coupling to intracellular second messengers, and distribution within the nervous system (Zifa and Fillion, 1992 [PubMed 1359584]). The serotonergic receptors belong to the multigene family of receptors coupled to guanine nucleotide-binding proteins.[supplied by OMIM]
FUNCTION: SwissProt: P28222 # 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. The activity of this receptor is mediated by G proteins that inhibit adenylate cyclase activity. SIZE: 390 amino acids; 43568 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. PTM: Phosphorylated. Desensitization of the receptor may be mediated by its phosphorylation. & Palmitoylated. SIMILARITY: SwissProt: P28222 ## Belongs to the G-protein coupled receptor 1 family.
Incubation Conditions
RECOMMENDED ASSAY 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 50mM Tris, 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 8-fold signal:background with 3H labeled GR125743 at 7 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst5 Receptor membrane prep.
10 µg/well
20 µg/well
Signal:Background
12.4
10.5
Specific Binding (cpm)
3210
3352
SPECIFICATIONS: 1 unit =10 μg Bmax for [3H]-GR125743 binding: 3.7 pmol/mg protein Kd for [3H]-GR125743 binding: ~10.5 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 Conditions
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size
200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Bestellnummer
GTIN
HTS108M
04053252510502
Documentation
ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor SDB
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 >>