Ordering Information
Product Images
Figure 1. Saturation binding for β3 receptor. 5 ug/well β3 Adrenoceptor Membrane Preparation was incubated with increasing amo ...
Figure 2. Competition binding for β3 adrenoceptor. β3 adrenoceptor Membrane Preparation (10µg/well) or Wild-Type Chem-1 membra ...
ChemiSCREEN™ Membrane Preparation Recombinant Human β3 Adrenoceptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human β3 Adrenoceptor
Trade Name:
Chemicon (Millipore)
Product 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.
Application Notes:
Radioligand binding assay, and GTPγS binding.
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.
Cell Membrane Type:
GPCR
Alternate Names:
beta3
Descriptive Text:
Host Cells:
Chem-1, a adherent mammalian cell line without any endogenous β3 adrenoceptor expression
Cell Type:
Chem-1
Gene Symbol:
- ADRB3
- ADRB3R
- B3AR
- BETA3AR
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°C.
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 Assurance:
| 10 µg/well | |
|---|---|
| Signal:Background | 13.3 |
| Specific Binding (cpm) | 11074.7 |
Bmax: 1.3 pmol/mg
Kd: ~0.69 nM
Protein or Enzyme Type:
GPCR Membrane Preparations
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.
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.
Storage Conditions:
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Species:
Human