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ChemiScreen™ Membrane Preparation Human Recombinant b3

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货号：HTS159M

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 beta 3
Description
Catalogue Number HTS159M
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiSCREEN™ Membrane Preparation Recombinant Human β3 Adrenoceptor
Overview Full-length human ADRB3 cDNA encoding the β3 adrenoceptor
Background Information The beta adrenergic receptors mediate the effects of endogenous catecholamines, such as epinephrine, by coupling to Gs to stimulate cAMP . Whereas β1 and β2 are found predominantly in heart, the β3 receptor is found primarily in adipose tissue. Activation of adipose β3 results in lipolysis and thermogenesis. A polymorphism in the human gene for β3 is associated with weight gain in obese patients (Clement et al., 1995). In addition, mice lacking the β3-adrenoceptor display increased total body fat, particularly on a high fat diet (Revelli et al., 1997). These observations indicate that β3 is a possible target for obesity treatments. Millipore's β3 adrenoceptor 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 antagonists of β3 adrenoceptor interactions. The membrane preparations exhibit a Kd of 0.69 nM for [125I]-(-) Iodocyanopindolol (ICYP). With 10 µg/well β;3 Adrenoceptor Membrane Prep and 0.75 nM [125I]-(-)ICYP, a greater than 8-fold signal-to-background ratio was obtained.
References
Product Information
Format Membranes
Presentation One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 8-fold signal:background with 125I-labeled (-)Iodocyanopindolol at 0.75 nM.

Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80&degC.
Applications
Application Human beta 3 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application Notes Radioligand binding assay, and GTPγS binding.
Biological Information
Entrez Gene Number
Entrez Gene Summary The ADRB3 gene product, beta-3-adrenergic receptor, is located mainly in adipose tissue and is involved in the regulation of lipolysis and thermogenesis. Beta adrenergic receptors are involved in the epenephrine and norepinephrine-induced activation of adenylate cyclase through the action of G proteins.
Gene Symbol
  • ADRB3
  • ADRB3R
  • B3AR
  • BETA3AR
GPCR Class A
Host Cells Chem-1
Protein Target beta 3
Target Sub-Family Adrenergic
UniProt Number
UniProt Summary FUNCTION: SwissProt: P13945 # Beta-adrenergic receptors mediate the catecholamine- induced activation of adenylate cyclase through the action of G proteins. Beta-3 is involved in the regulation of lipolysis and thermogenesis.
SIZE: 408 amino acids; 43519 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed mainly in adipose tissues.
SIMILARITY: SwissProt: P13945 ## 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. 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, filtered and stored at 4°C
Radioligand: [125I]-(-) Iodocyanopindolol (Perkin Elmer # NEX189)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, filtered and stored at 4°C.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
10 µg/well
Signal:Background 13.3
Specific Binding (cpm) 11074.7
SPECIFICATIONS: 1 unit = 10 µg membrane preparation
Bmax: 1.3 pmol/mg
Kd: ~0.69 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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