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HTS108M ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor

Human 5-HT1B 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: P28222 Entrez Gene Number: NM_000863.1
HTS108M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-15-HT1B
Description
Catalogue NumberHTS108M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor
OverviewFull-length human HTR1B cDNA encoding the 5-HT1B Serotonin Receptor
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). 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
FormatMembranes
PresentationLiquid 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.
Quality LevelMQ100
Applications
ApplicationHuman 5-HT1B 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 SummaryThe 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]
Gene Symbol
  • HTR1B
  • S12
  • 5-HT1B
  • HTR1D2
  • HTR1DB
  • 5-HT1DB
  • 5-HT-1D-beta
  • 5-HT-1B
GPCR ClassA
Host CellsChem-1
Protein Target5-HT1B
Target Sub-FamilySerotonin
UniProt Number
UniProt SummaryFUNCTION: 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 ConditionsRECOMMENDED 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

Radioligand: [3H]-GR125743. (Amersham #:TRK-1046 )

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
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst5 Receptor membrane prep.
Product Usage Statements
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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS108M 04053252510502

Documentation

ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor SDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT1B Serotonin Receptor Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT1B SEROTONIN RECEPTOR - 2341138 2341138
CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 5-HT1B SEROTONIN RECEPTOR - 2250864 2250864

Data Sheet

Title
CHEMISCREENTM MEMBRANE PREPARATION HUMAN RECOMBINANT 5-HT1B SEROTONIN RECEPTOR

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