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HTS096M ChemiSCREEN™ Human α2A Adrenergic Receptor Membrane Preparation

Human alpha2A GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) o SDS, certificato d’analisi (CoA) e certificato di qualità (CoQ), dossier, brochure e altri documenti disponibili.
Host Cell: Chem-1 Uni Prot Number: P08913 Entrez Gene Number: NM_000681.2
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HTS096M
200 units  
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      Tabella delle specifiche principali

      SpeciesHost CellsProtein Target
      Human Chem-1alpha2A
      Description
      Catalogue NumberHTS096M
      Brand Family Chemicon®
      Trade Name
      • ChemiScreen
      • Chemicon
      DescriptionChemiSCREEN™ Human α2A Adrenergic Receptor Membrane Preparation
      OverviewFull-length human ADRA2A transcript variant 1 cDNA encoding α2A
      Background InformationThe 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 α2A 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 α2A. The membrane preparations exhibit a Kd of 5.4 nM for [3H]-MK-912. With 5 nM [3H]-MK-912, 5 µg/well α2A Membrane Prep typically yields greater than 20-fold signal-to-background ratio.
      References
      Product Information
      FormatMembranes
      PresentationOne 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 MK-912 at 5 nM

      Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
      Quality LevelMQ100
      Applications
      ApplicationHuman alpha2A GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
      Key Applications
      • Radioligand binding assay
      • GTPγS binding
      Application NotesRadioligand binding assay and GTPγS binding.
      Biological Information
      Entrez Gene Number
      Entrez Gene SummaryAlpha-2-adrenergic receptors are members of the G protein-coupled receptor superfamily. They include 3 highly homologous subtypes: alpha2A, alpha2B, and alpha2C. These receptors have a critical role in regulating neurotransmitter release from sympathetic nerves and from adrenergic neurons in the central nervous system. Studies in mouse revealed that both the alpha2A and alpha2C subtypes were required for normal presynaptic control of transmitter release from sympathetic nerves in the heart and from central noradrenergic neurons; the alpha2A subtype inhibited transmitter release at high stimulation frequencies, whereas the alpha2C subtype modulated neurotransmission at lower levels of nerve activity. This gene encodes alpha2A subtype and it contains no introns in either its coding or untranslated sequences.
      Gene Symbol
      • ADRA2A
      • ZNF32
      • ADRA2R
      • ALPHA2AAR
      • Alpha-2AAR
      • ADRA2
      • OTTHUMP00000058849
      • ADRAR
      GPCR ClassA
      Host CellsChem-1
      Protein Targetalpha2A
      Target Sub-FamilyAdrenergic
      UniProt Number
      UniProt SummaryFUNCTION: SwissProt: P08913 # Alpha-2 adrenergic receptors mediate the catecholamine- induced inhibition of adenylate cyclase through the action of G proteins. The rank order of potency for agonists of this receptor is oxymetazoline > clonidine > epinephrine > norepinephrine > phenylephrine > dopamine > p-synephrine > p-tyramine > serotonin = p-octopamine. For antagonists, the rank order is yohimbine > phentolamine = mianserine > chlorpromazine = spiperone = prazosin > propanolol > alprenolol = pindolol.
      SIZE: 450 amino acids; 48957 Da
      SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
      SIMILARITY: SwissProt: P08913 ## Belongs to the G-protein coupled receptor 1 family.
      Incubation ConditionsMembranes 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] MK-912 (Perkin Elmer#: NET-1059)
      Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Table 1. Signal:background and specific binding values obtained in a competition binding assay with α2A membrane prep and unlabeled rauwolscine.
      Product Usage Statements
      10 µg/well 5 µg/well
      Signal:Background 28.97 34.14
      Specific Binding (cpm) 11777 9147



      SPECIFICATIONS: 1 unit = 5 µg
      Bmax for [3H] MK-912 binding: 71.4 pmol/mg protein
      Kd for [3H] MK-912 binding: ~5.4 nM 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 ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
      Packaging Information
      Material Size200 units
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Numero di catalogo GTIN
      HTS096M 04053252334894

      Documentation

      ChemiSCREEN™ Human α2A Adrenergic Receptor Membrane Preparation MSDS

      Titolo

      Scheda di sicurezza (MSDS) 

      ChemiSCREEN™ Human α2A Adrenergic Receptor Membrane Preparation Certificati d'Analisi

      TitoloNumero di lotto
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN #945;2A ADRENERGIC RECEPTOR - 2091385 2091385

      Riferimenti bibliografici

      Panoramica dei riferimenti bibliograficiCodice d'identificazione nel Pub Med
      In vivo gene modification elucidates subtype-specific functions of alpha(2)-adrenergic receptors
      Kable, J W, et al
      J Pharmacol Exp Ther, 293:1-7 (2000)  1999

      		10734146 10734146

      Scheda tecnica

      Titolo
      CHEMISCREENTM MEMBRANE PREPARATION RECOMBINANT HUMAN 2A ADRENERGIC RECEPTOR

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      Categorie

      Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
      Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations