Ordering Information
Product Images
Figure 1. Saturation binding for D3 Receptor. 5.0 µg/well D3 Membrane Preparation was incubated with increasing amount of [125I]-Iodospiperone in th ...
Figure 2. Competition binding for D3 Receptor. D3 Receptor Membrane Preparation (5.0 or 10µg/well) or Wild-Type Chem-1 membrane preparation (WT; Ch ...
ChemiSCREEN™ Human D3 Dopamine Receptor Membrane Preparation
Description:
ChemiSCREEN™ Human D3 Dopamine Receptor Membrane Preparation
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Product Overview:
Human D3 Receptor
Background Information:
Dopamine is a catecholamine neurotransmitter that functions in the CNS to control locomotor, cognitive, emotional and neurendocrine processes, and in the periphery to modulate cardiovascular, renal and gastrointestinal processes. The biological activities of dopamine are mediated by a family of five GPCRs. The D1 and D5 subtypes couple to Gs to increase intracellular cAMP, whereas the D2, D3 and D4 subtypes couple to Gi to reduce cAMP (Missale et al., 1998). The dopamine D3 receptor is expressed in phylogenetically older regions of the brain, suggesting that it plays a role in cognitive and emotional functions. It has been the target of potential drugs for psychotic disorders and Parkinson's disease. Chemicon's D3 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 D3 interactions and its ligands. The membrane preparations exhibit a Kd of 0.1 nM for [125I]-Iodospiperone. With 5.0ug/well D3 membrane prep and 0.1 nM [125I]-Iodospiperone, a greater than 4-fold signal-to-background ratio was obtained.
Application Notes:
Radioligand binding assay and GTPγS binding
Species Reactivity:
Human
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:
DRD3
Descriptive Text:
Host Cells:
Chem-1, an adherent mammalian cell line without any endogenous D3 Receptor expression.
Cell Type:
Chem-1
Gene Symbol:
- DRD3
- FET1
- MGC149204
- D3DR
- ETM1
- D
- MGC149205
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 4-fold signal:background with 125I-labeled Iodospiperone at 0.1 nM.
Liquid in packaging buffer: 50 mM Tris, pH 7.4, 10% glycerol with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Liquid in packaging buffer: 50 mM Tris, pH 7.4, 10% glycerol with no preservatives.
Packaging method: Membranes protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.
Quality Assurance:
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of D3 Receptor membrane prep.
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 0.32 pmol/mg
Kd: 0.1 nM
| 10 µg/well | 5 µg/well | |
|---|---|---|
| Signal:Background | 4.88 | 5.93 |
| Specific Binding (cpm) | 1853.7 | 1861.3 |
SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 0.32 pmol/mg
Kd: 0.1 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, a GF/C 96-well filter plate 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, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] -Iodospiperone(Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
Binding buffer: 50 mM Tris, pH 7.4, 120mM NaCl, filtered and stored at 4°C
Radioligand: [125I] -Iodospiperone(Perkin Elmer # NEX284)
Wash Buffer: 50 mM Tris, pH 7.4, filtered and stored at 4°C.
Species:
Human
Storage Conditions:
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.