HTS020M
ChemiSCREEN Membrane Preparation Recombinant Human CB2 Cannabinoid Receptor
Human CB2 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Sinonimo/i:
Cannabinoid receptor membrane prep, Human CB2 membrane prep
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About This Item
Codice UNSPSC:
41106514
eCl@ss:
32161000
NACRES:
NA.41
Prodotti consigliati
Origine biologica
human
Livello qualitativo
Ricombinante
expressed in CHO 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
Descrizione generale
Cannabinoid compounds include exogenous drugs such as D9-THC, the main psychoactive component of the plant Cannabis sativa, and endogenous mediators, such as anandamide, that belong to eicosanoid family.The biological effects of cannabinoids are mediated by a family of two Gi-coupled 7-transmembrane receptors, CB1 and CB2. The CB1 receptor is found primarily in brain and mediates the psychoactive effects of cannabinoid ligands. The CB2 receptor is expressed mainly in immune cells, including mast cells and CD40-activated B cells, where it mediates proliferation and inhibition of migration (Howlett et al., 2002). Activation of CB2 inhibits the development of liver fibrosis (Julien et al., 2005). In bone, CB2 is expressed in both osteoblasts and osteoclasts, and functions to prevent bone loss (Ofek et al., 2006). In addition, activation of CB2 has an antinociceptive effect in animal models of neuropathic, inflammatory, and acute pain; this effect is mediated by release of endogenous opioids in the periphery (Ibrahim et al., 2005). Millipore′s CB2 membrane preparations are crude membrane preparations made from stable recombinant cell lines with a high-level of GPCR surface expression; thus, they are ideal HTS tools for screening for agonists and antagonists of CB2. The membrane preparations exhibit a Kd of 2.7 nM for [3H]-CP55,940. With 5 μg/well CB2 Membrane Prep and 2.5 nM [3H]-CP55,940, greater than 10-fold signal-to-background ratio is obtained.
Full-length Human CB2
Qualità
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of CB2 membrane prep.
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 50 pmol/mg
Kd: 2.7 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 33.5 | 25.2 |
| Specific Binding (cpm) | 7336 | 6207 |
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 50 pmol/mg
Kd: 2.7 nM
Specifiche
Inucbation Conditions
RECOMMENDED 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 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]-CP55940 (Perkin Elmer #NET1051)
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.
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 3H-labeled CP55,940 at 2.5 nM
RECOMMENDED 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 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]-CP55940 (Perkin Elmer #NET1051)
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.
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 3H-labeled CP55,940 at 2.5 nM
Stato fisico
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.
Packaging method: Membranes proteins were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Packaging method: Membranes proteins were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
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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Alexei Yeliseev et al.
Scientific reports, 11(1), 3706-3706 (2021-02-14)
Signaling through integral membrane G protein-coupled receptors (GPCRs) is influenced by lipid composition of cell membranes. By using novel high affinity ligands of human cannabinoid receptor CB2, we demonstrate that cholesterol increases basal activation levels of the receptor and alters
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