HTS215M
ChemiSCREEN Membrane Preparation Recombinant Human 5-HT₇ Serotonin Receptor
Human 5-HT7 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Codice UNSPSC:
41106514
eCl@ss:
32161000
NACRES:
NA.84
Prodotti consigliati
Origine biologica
human
Livello qualitativo
Ricombinante
expressed in Chem-1 cells
Produttore/marchio commerciale
ChemiScreen
Chemicon®
tecniche
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
N° accesso NCBI
N° accesso UniProt
Condizioni di spedizione
dry ice
Informazioni sul gene
human ... HTR7(3363)
Descrizione generale
Full-length human HTR7 cDNA encoding the 5-HT₇ Serotonin Receptor
The 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.
Applicazioni
Human 5-HT7 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Azioni biochim/fisiol
Protein Target: 5-HT7
Qualità
Table 1. Signal:background and specific binding values obtained in a competition binding assay with 10 μg of 5-HT₇ Receptor membrane prep.
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
| 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
Specifiche
Inucbation 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 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
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 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
Stato fisico
Liquid 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.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Stoccaggio e stabilità
Maintain frozen at –70°C for up to 2 years. Do not freeze and thaw.
Note legali
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Codice della classe di stoccaggio
12 - Non Combustible Liquids
Classe di pericolosità dell'acqua (WGK)
WGK 2
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
Certificati d'analisi (COA)
Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.
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I documenti relativi ai prodotti acquistati recentemente sono disponibili nell’Archivio dei documenti.
J A Terrón et al.
British journal of pharmacology, 127(3), 609-616 (1999-07-13)
1. We investigated in the present study whether 5-HT is able to exert direct relaxant responses in canine basilar and middle cerebral arteries via the 5-HT7 receptor. 2. In arterial rings deprived of endothelium and pre-contracted with prostaglandin F2 alpha
U L Mullins et al.
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology, 21(3), 352-367 (1999-08-24)
Recent evidence suggests that a novel serotonin receptor 5-HT7 localized in the hypothalamus downregulates in response to treatment with the antidepressant fluoxetine (Sleight et al. 1995). This receptor has also been implicated in the regulation of circadian rhythms (Lovenberg et
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