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HTS104M ChemiSCREEN™ Membrane Preparation Recombinant Human β1 Adrenoceptor Receptor

Human beta 1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P08588 Entrez Gene Number: NM_000684.2
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HTS104M
200 units  
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Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1beta 1
Description
Catalogue NumberHTS104M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human β1 Adrenoceptor Receptor
OverviewFull-length human ADRB1 cDNA encoding the β1 adrenoceptor
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-adrenergic receptors (Bylund et al., 1994). The three members of the β-adrenergic receptor family, β1, β2 and β3, couple to Gs to increase cAMP upon activation. In the heart, the β1 receptor constitutes 70-80% of the β-adrenergic receptors. Activation of cardiac β-adrenergic receptors acutely increases heart rate, cardiac output, and cardiac automaticity, and chronically increases cardiac myocyte apoptosis. In failing hearts, the β1 subtype is downregulated and desensitized, probably as a result of increased catecholamine levels. As a result, β-adrenergic receptor antagonists (β blockers) are effective in the treatment of congestive heart failure and arrhythmia (Lohse et al., 2003). Millipore's β1 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 β1 adrenoceptor interactions. The membrane preparations exhibit a Kd of 2.6 nM for [125I]-(-) Iodocyanopindolol (ICYP). With 5 µg/well β1 Adrenoceptor Membrane Prep and 0.25 nM [125I]-(-)ICYP, a greater than 15-fold signal-to-background ratio was obtained.
References
Product Information
FormatMembranes
PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 15-fold signal:background with 125I-labeled (-)Iodocyanopindolol at 0.25 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°C.
Quality LevelMQ100
Applications
ApplicationHuman beta 1 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 SummaryThe adrenergic receptors (subtypes alpha 1, alpha 2, beta 1, and beta 2) are a prototypic family of guanine nucleotide binding regulatory protein-coupled receptors that mediate the physiological effects of the hormone epinephrine and the neurotransmitter norepinephrine. Specific polymorphisms in this gene have been shown to affect the resting heart rate and can be involved in heart failure.
Gene Symbol
  • ADRB1
  • ADRB1R
  • RHR
  • B1AR
  • BETA1AR
GPCR ClassA
Host CellsChem-1
Protein Targetbeta 1
Target Sub-FamilyAdrenergic
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P08588 # Beta-adrenergic receptors mediate the catecholamine- induced activation of adenylate cyclase through the action of G proteins. This receptor binds epinephrine and norepinephrine with approximately equal affinity.
SIZE: 477 amino acids; 51323 Da
SUBUNIT: Interacts with GOPC and DLG4.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. Note=Localized at the plasma membrane. Found in the Golgi upon GOPC overexpression.
DOMAIN: SwissProt: P08588 The PDZ domain-binding motif mediates competitive interactions with GOPC and DLG4 and plays a role in subcellular location of the receptor.
PTM: Homologous desensitization of the receptor is mediated by its phosphorylation by beta-adrenergic receptor kinase.
SIMILARITY: 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. 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
5 µg/well 10 µg/well
Signal:Background 28.5 34.6
Specific Binding (cpm) 27002 28467


SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 47.9 pmol/mg
Kd: ~2.6 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
Catalogue Number GTIN
HTS104M 04053252285608

Documentation

ChemiSCREEN™ Membrane Preparation Recombinant Human β1 Adrenoceptor Receptor SDS

Title

Safety Data Sheet (SDS) 

References

Reference overviewPub Med ID
What is the role of beta-adrenergic signaling in heart failure?
Lohse, Martin J, et al.
Circ. Res., 93: 896-906 (2003)  2003

Show Abstract 		14615493 14615493

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