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Die folgenden MAPmates™ sollten nicht zusammen analysiert werden: -MAPmates™, die einen unterschiedlichen Assaypuffer erfordern. -Phosphospezifische und MAPmate™ Gesamtkombinationen wie Gesamt-GSK3β und Gesamt-GSK3β (Ser 9). -PanTyr und locusspezifische MAPmates™, z.B. Phospho-EGF-Rezeptor und Phospho-STAT1 (Tyr701). -Mehr als 1 Phospho-MAPmate™ für ein einziges Target (Akt, STAT3). -GAPDH und β-Tubulin können nicht mit Kits oder MAPmates™, die panTyr enthalten, analysiert werden.
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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
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CB1 is a GPCR that is expressed primarily in brain and nervous tissue, and mediates numerous CNS responses such as analgesia, appetite, cognition, memory and locomotor activity. A number of cannabinoid ligands bind to CB1 and activate Gi/o-mediated downstream responses, including inhibition of cAMP production and activation of ion channels and MAP kinases. Ligands for CB1 include exogenous agonists such as Delta9-THC, the main psychoactive component of the plant Cannabis sativa, and endogenous eicosanoid agonists such as anandamide. A number of synthetic agonists such as CP55940 and R-(+)-WIN55212, and antagonists, such as SR141716A, for CB1 have been developed (Howlett et al., 2002). CB1 agonists have clinical utility in analgesia and antiemetic properties, whereas CB1 antagonists show promise for treatment of appetite in obesity disorders. Chemicon's CB1 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 for agonists and antagonists at CB1. The membrane preparations exhibit a Kd of 7 nM for [3H]-SR141716A. In the presence of 2 nM [3H]-SR141716A with CP55940 as unlabeled competitor, 5 μg/well and 10 μg/well CB1 Membrane Prep yield 4- and 5-fold signal-to-background ratios, respectively.
References
Product Information
Format
Membranes
Presentation
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
This gene encodes a protein that is one of two cannabinoid receptors. The cannabinoids, principally delta-9-tetrahydrocannabinol and synthetic analogs, are psychoactive ingredients of marijuana. The cannabinoid receptors are members of the family of guanine-nucleotide-binding protein (G-protein) coupled receptors which inhibit adenylate cyclase activity in a dose-dependent, stereoselective and pertussis toxin-sensitive manner. The two receptors have been found to be involved in the cannabinoid-induced CNS effects (including alterations in mood and cognition) experienced by users of marijuana. Two transcript variants encoding different isoforms have been described for this gene.
FUNCTION: SwissProt: P21554 # Involved in cannabinoid-induced CNS effects. Acts by inhibiting adenylate cyclase. Could be a receptor for anandamide. Isoform 2 and isoform 3 have altered ligand binding. SIZE: 472 amino acids; 52858 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. TISSUE SPECIFICITY: Widely expressed. SIMILARITY: SwissProt: P21554 ## Belongs to the G-protein coupled receptor 1 family.
Incubation 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, a GF/C 96-well filter plate 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. Ligands were diluted in binding buffer containing 30% DMSO, then added to membranes such that the final DMSO concentration was 15%.Radioligand: [3H] SR141716A (Amersham #TRK1028)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
One vial contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield approximately 4-fold signal:background with [3H] SR141716A at 2 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance
SPECIFICATIONS: 1 unit = 10 μg Bmax: 15 pmol/mg Kd: 7 nM for SR141716A
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 Conditions
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Millipore offers a large selection of robust and reliable G-protein coupled receptor products, including Stable Cell Lines, Membrane Preparations, and Frozen Cells. See below for GPCR research tools. Weitere Informationen >>
