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96-Well Plate
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48-602MAG
Buffer Detection Kit for Magnetic Beads
1 Kit
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HTS158M
Sigma-AldrichChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor
Human alpha1B GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
ChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor
Overview
Full-length human ADRA1B cDNA encoding α1B adrenergic
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 three members of the α1 subclass of adrenoceptors, α1A, α1B and α1D, couple to Gq, and promote contraction of vascular and urinary tract smooth muscle, relaxation of intestinal smooth muscle, increased contractile force in the heart, and glycogenolysis and gluconeogenesis in the liver. The different subtypes have overlapping distributions and variably contribute to these effects depending on species and tissue. Overexpression of a constitutively active α1B mutant in the heart of transgenic mice resulted in cardiac hypertrophy with increased heart weight/body weight ratios. Analysis of α1B knock out mice has provided evidence that α1B is a mediator of blood pressure and aortic contractile responses induced by α1 agonists (Milano et al., 1994). The locomotor and rewarding effects of pysochostimulants and opiates were suppressed in mice lacking α1B-adrenergic receptors (Drouin et al. 2002). Millipore's α1B 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 of α1B. The membrane preparations exhibit a Kd of 0.8 nM for [3H]-Prazosin. With 1 nM [3H]-Prazosin, 5μg/well α1B Membrane Prep typically yields greater than 5-fold signal-to-background ratio.
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 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C
Alpha-1-adrenergic receptors (alpha-1-ARs) are members of the G protein-coupled receptor superfamily. They activate mitogenic responses and regulate growth and proliferation of many cells. There are 3 alpha-1-AR subtypes: alpha-1A, -1B and -1D, all of which signal through the Gq/11 family of G-proteins and different subtypes show different patterns of activation. This gene encodes alpha-1B-adrenergic receptor, which induces neoplastic transformation when transfected into NIH 3T3 fibroblasts and other cell lines. Thus, this normal cellular gene is identified as a protooncogene. This gene comprises 2 exons and a single large intron of at least 20 kb that interrupts the coding region.
FUNCTION: SwissProt: P35368 # This alpha-adrenergic receptor mediates its action by association with G proteins that activate a phosphatidylinositol- calcium second messenger system. SIZE: 520 amino acids; 56836 Da SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein. SIMILARITY: SwissProt: P35368 ## 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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM Tris, 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 Tris, pH 7.4, 10 mM MgCl2, 1 mM EDTA, filtered and stored at 4°C
Wash Buffer: 50 mM Tris, pH 7.4, 500mM NaCl . 0.1% BSA filtered and stored at 4°C.
One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 5-fold signal:background with 3H labeled Prazosin at 1nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of α1B Receptor membrane prep.
5 µg/well
Signal:Background
11.8
Specific Binding (cpm)
1171
SPECIFICATIONS: 1 unit = 5 µg membrane preparation Bmax for [3H]-Prazosin binding: 12.9 pmol/mg protein Kd ffor [3H]-Prazosin binding: ~0.8 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
Global Trade Item Number
Bestellnummer
GTIN
HTS158M
04053252328619
Documentation
ChemiSCREEN™ Membrane Preparation Recombinant Human α1B Adrenergic Receptor SDB
Decreased blood pressure response in mice deficient of the alpha1b-adrenergic receptor. Cavalli, A, et al. Proc. Natl. Acad. Sci. U.S.A., 94: 11589-94 (1997)
1997
Myocardial expression of a constitutively active alpha 1B-adrenergic receptor in transgenic mice induces cardiac hypertrophy. Milano, C A, et al. Proc. Natl. Acad. Sci. U.S.A., 91: 10109-13 (1994)
1993
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 >>
