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HTS215M ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor

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HTS215M
200 units  
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Overview

Replacement Information

Key Specifications Table

SpeciesHost CellsProtein Target
Human Chem-15-HT7
Description
Catalogue NumberHTS215M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor
OverviewFull-length human HTR7 cDNA encoding the 5-HT₇ Serotonin Receptor
Background InformationThe neurotransmitter serotonin/5-hydroxytryptamine (5-HT) regulates a wide variety of neurological functions. A family of 13 receptors (12 GPCRs and one ion channel) mediate the effects of serotonin. The serotonin receptor 5-HT₇ is a Gs coupled receptor expressed in the brain (hypothalamus, hippocampus and frontal cortex) and in smooth muscle and GI tract. Systemic delivery of serotonin results in hypothermia, and pharmacological and genetic studies indicate that 5-HT₇ mediates this effect (Hedlund et al., 2003). In addition, 5-HT₇ appears to modulate circadian rhythms and smooth muscle relaxation, and is implicated as a target for sleep, particularly as related to depression, and migraine (Terron and Falcon-Neri, 1999; Mullins et al., 1999). Millipore's 5-HT₇ 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-HT₇. The membrane preparations exhibit a Kd of 37.7 nM for [³H]-5-Carboxamidotryptamine (5-CT). With 30 nM [³H]-5-CT, 10 μg/well 5-HT₇ Membrane Prep typically yields greater than 4-fold signal-to-background ratio.
References
Product Information
FormatMembranes
HS Code3822 19 90
PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman 5-HT7 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
Gene Symbol
  • HTR7
GPCR ClassA
Host CellsChem-1
Protein Target5-HT7
Target Sub-FamilySerotonin
UniProt Number
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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with the binding buffer. 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, 5 mM MgCl₂, 4 mM CaCl₂, 0.5 mM ascorbic acid, filtered and stored at 4°C

Radioligand: [³H]-5-Carboxamidotryptamine (Perkin Elmer NET1071)

Wash Buffer: same as the binding buffer.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 4-fold signal:background with ³H-labeled 5-CT at 30 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 10 μg of 5-HT₇ Receptor membrane prep.
Product Usage Statements
10 µg/well
Signal:Background 7.5
Specific Binding (cpm) 545

SPECIFICATIONS: 1 unit =10 μg
Bmax for [³H]-5-CT binding: 20.1 pmol/mg protein
Kd for [³H]-5-CT binding: ~37.7 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
Catalog Number GTIN
HTS215M 04053252474309

Documentation

ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor SDS

Title

Safety Data Sheet (SDS) 

ChemiSCREEN™ Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor Certificates of Analysis

TitleLot Number
CHEMISCREENTM MEMBRANE PREPARATION HUMAN RECOMBINANT 5-HT7 SEROTONIN RECEPTOR - 2324476 2324476

References

Reference overviewPub Med ID
No hypothermic response to serotonin in 5-HT7 receptor knockout mice.
Hedlund, P B, et al.
Proc. Natl. Acad. Sci. U.S.A., 100: 1375-80 (2003)  2003

12529502 12529502
Effects of antidepressants on 5-HT7 receptor regulation in the rat hypothalamus.
Mullins, U L, et al.
Neuropsychopharmacology, 21: 352-67 (1999)  1999

10457532 10457532
Pharmacological evidence for the 5-HT7 receptor mediating smooth muscle relaxation in canine cerebral arteries.
Terrón, J A and Falcón-Neri, A
Br. J. Pharmacol., 127: 609-16 (1999)  1999

10401550 10401550

Data Sheet

Title
CHEMISCREEN™ MEMBRANE PREPARATION HUMAN RECOMBINANT 5-HT7 SEROTONIN RECEPTOR

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Categories

Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations