HTS157M
ChemiSCREEN Human α2B Adrenergic Receptor Membrane Preparation
Human alpha2B GPCR membrane prepation for Radioligand binding Assays.
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About This Item
Codice UNSPSC:
41106514
eCl@ss:
32161000
NACRES:
NA.84
Prezzi e disponibilità al momento non sono disponibili
Prodotti consigliati
Origine biologica
human
Livello qualitativo
Ricombinante
expressed in Chem-1 cells
Produttore/marchio commerciale
ChemiScreen
Chemicon®
tecniche
radioligand binding assay (RLBA): suitable
N° accesso NCBI
N° accesso UniProt
Condizioni di spedizione
dry ice
Descrizione generale
Full-length human ADRA2B transcript cDNA encoding α2B.
The endogenous catecholamines epinephrine and norepinephrine have profound effects on smooth muscle activity, cardiac function, carbohydrate and fat metabolism, hormone secretion, neurotransmitter release, and central nervous system actions. These activities are mediated by GPCRs belonging to two subfamilies, the α- and β-adrenoceptors (Bylund et al., 1994). The α2 adrenergic receptor subfamily members, consisting of α2A, α2B, and α2C, couple primarily to Gi to inhibit cAMP production, and play an important role in regulation of cardiovascular and CNS function. Experiments with α2A-selective agonists and mice lacking α2A demonstrate that activation of α2A results in hypotension, sedation, analgesia, and hypothermia (Kable et al., 2000). Chemicon′s α2B 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 at α2B. The membrane preparations exhibit a Kd of 11.5 nM for [3H]-Rauwolscine. With 15 nM [3H]-Rauwolscine, 5 µg/well α2B Membrane Prep typically yields greater than 30-fold signal-to-background ratio.
Azioni biochim/fisiol
Protein Target: alpha2B
Qualità
Table 1. Signal:background and specific binding values obtained in a competition binding assay with α2B membrane prep and unlabeled rauwolscine.
SPECIFICATIONS: 1 unit = 5 µg
Bmax for [3H]-rauwolscine binding: 154 pmol/mg protein
Kd for [3H]-rauwolscine binding: ~11.5 nM
| 5 µg/well | |
|---|---|
| Signal:Background | 64.7 |
| Specific Binding (cpm) | 14646 |
SPECIFICATIONS: 1 unit = 5 µg
Bmax for [3H]-rauwolscine binding: 154 pmol/mg protein
Kd for [3H]-rauwolscine binding: ~11.5 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 50mM HEPES, pH 7.4, 0.5% BSA. 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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [3H]-Rauwolscine (Perkin Elmer#: NET-722)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, 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 HEPES, pH 7.4, 0.5% BSA. 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 Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [3H]-Rauwolscine (Perkin Elmer#: NET-722)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
Stato fisico
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 20-fold signal:background with 3H-labeled Rauwolscine at 15 nM
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
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)
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I documenti relativi ai prodotti acquistati recentemente sono disponibili nell’Archivio dei documenti.
J W Kable et al.
The Journal of pharmacology and experimental therapeutics, 293(1), 1-7 (2000-03-29)
Mice with altered alpha(2)-adrenergic receptor genes have become important tools in elucidating the subtype-specific functions of the three alpha(2)-adrenergic receptor subtypes because of the lack of sufficiently subtype-selective pharmacological agents. Mice with a deletion (knockout) of the alpha(2A)-, alpha(2B)-, or
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