HTS039M
ChemiSCREEN Membrane Preparation Recombinant Human D2 Dopamine Receptor
Human D2L GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
Kod UNSPSC:
41106514
eCl@ss:
32161000
NACRES:
NA.84
Polecane produkty
pochodzenie biologiczne
human
Poziom jakości
rekombinowane
expressed in Chem-1 cells
producent / nazwa handlowa
ChemiScreen
Chemicon®
metody
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
numer dostępu NCBI
numer dostępu UniProt
Warunki transportu
dry ice
Opis ogólny
Dopamine is a catecholamine neurotransmitter that functions in the CNS to control locomotor, cognitive, emotional and neurendocrine processes, and in the periphery to modulate cardiovascular, renal and gastrointestinal processes. The biological activities of dopamine are mediated by a family of five GPCRs. The D1 and D5 subtypes couple to Gs to increase intracellular cAMP, whereas the D2, D3 and D4 subtypes couple to Gi to reduce cAMP (Missale et al., 1998). The D2 dopamine receptors have been of particular clinical interest due to their regulation of prolactin secretion and their affinity for antipsychotic drugs. The D2 receptor exists as two alternatively spliced isoforms differing in the insertion of a stretch of 29 amino acids in the third intracellular loop (D2S and D2L) (Giros et al., 1989; Grandy et al., 1989). Millipore′s D2 dopamine 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 D2 dopamine receptor interactions with its ligand. The membrane preparations exhibit a Kd of 0.51nM for [125I]-Iodospiperone. With 5.0 ug/well D2 Dopamine Membrane Prep and 0.1nM [125I]-Iodospiperone, a greater than 10-fold signal-to-background ratio was obtained.
Human Dopamine Receptor 2
Zastosowanie
Radioligand binding assay and GTPγS binding
Działania biochem./fizjol.
GPCR Class: A
Protein Target: D2L
Target Sub-Family: Dopamine
Jakość
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of Dopamine Receptor membrane prep.
SPECIFICATIONS: 1 unit = 5.0 µg membrane preparation
Bmax: 6.03 pmol/mg
Kd: 0.51 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 23.6 | 27.5 |
| Specific Binding (cpm) | 8331 | 9927.2 |
Bmax: 6.03 pmol/mg
Kd: 0.51 nM
Charakterystyka techniczna
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 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, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] Iodospiperone (Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
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, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] Iodospiperone (Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
Postać fizyczna
One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 10-fold signal:background with [125I]-labeled Iodospiperone at 0.1 nM.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Przechowywanie i stabilność
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Informacje prawne
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Oświadczenie o zrzeczeniu się odpowiedzialności
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.
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Kod klasy składowania
12 - Non Combustible Liquids
Klasa zagrożenia wodnego (WGK)
WGK 2
Temperatura zapłonu (°F)
Not applicable
Temperatura zapłonu (°C)
Not applicable
Certyfikaty analizy (CoA)
Poszukaj Certyfikaty analizy (CoA), wpisując numer partii/serii produktów. Numery serii i partii można znaleźć na etykiecie produktu po słowach „seria” lub „partia”.
Masz już ten produkt?
Dokumenty związane z niedawno zakupionymi produktami zostały zamieszczone w Bibliotece dokumentów.
C Missale et al.
Physiological reviews, 78(1), 189-225 (1998-02-11)
The diverse physiological actions of dopamine are mediated by at least five distinct G protein-coupled receptor subtypes. Two D1-like receptor subtypes (D1 and D5) couple to the G protein Gs and activate adenylyl cyclase. The other receptor subtypes belong to
B Giros et al.
Nature, 342(6252), 923-926 (1989-12-21)
Dopamine receptors are classified into D1 and D2 subtypes on the basis of their pharmacological properties and the intracellular responses they mediate. The cerebral D2 dopamine receptor is the target of drugs used to alleviate the main symptoms of schizophrenia.
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