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ChemiScreen™ Membrane Prep Human Recombinant a2B

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货号：HTS157M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 alpha2B
Description
Catalogue Number HTS157M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ Human α2B Adrenergic Receptor Membrane Preparation
Overview Full-length human ADRA2B transcript cDNA encoding α2B.
Background Information 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.
References
Product Information
Format Membranes
Presentation 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.
Applications
Application Human alpha2B GPCR membrane prepation for Radioligand binding Assays.
Key Applications
  • Radioligand binding assay
Biological Information
Entrez Gene Number
Entrez Gene Summary Alpha-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. This gene encodes the alpha2B subtype, which was observed to associate with eIF-2B, a guanine nucleotide exchange protein that functions in regulation of translation. A polymorphic variant of the alpha2B subtype, which lacks 3 glutamic acids from a glutamic acid repeat element, was identified to have decreased G protein-coupled receptor kinase-mediated phosphorylation and desensitization; this polymorphic form is also associated with reduced basal metabolic rate in obese subjects and may therefore contribute to the pathogenesis of obesity. This gene contains no introns in either its coding or untranslated sequences.
Gene Symbol
  • ADRA2B
  • ADRA2L1
  • ALPHA2BAR
  • ADRA2RL1
  • ADRARL1
GPCR Class A
Host Cells Chem-1
Protein Target alpha2B
Target Sub-Family Adrenergic
UniProt Number
UniProt Summary FUNCTION: SwissProt: P18089 # 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 clonidine > norepinephrine > epinephrine = oxymetazoline > dopamine > p-tyramine = phenylephrine > serotonin > p-synephrine / p-octopamine. For antagonists, the rank order is yohimbine > chlorpromazine > phentolamine > mianserine > spiperone > prazosin > alprenolol > propanolol > pindolol.
SIZE: 450 amino acids; 49954 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P18089 ## Belongs to the G-protein coupled receptor 1 family.
Incubation 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.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
2B Table 1. Signal:background and specific binding values obtained in a competition binding assay with α membrane prep and unlabeled rauwolscine.
Product Usage Statements
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 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.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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