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Competition binding for sst1. sst1 Membrane Preparation (5 μg/well) and Wild-Type Chem-1 membrane preparation (10 g/well) were incubated with 0.5 nM...
Saturation binding for sst1. 5 ug/well sst1 Membrane Preparation was incubated with increasing amount of [125I]-somatostatin-14 in the abs...
ChemiSCREEN™ Membrane Preparation Recombinant Human sst1 Somatostatin Receptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human sst1 Somatostatin Receptor
Trade Name:
- Chemicon (Millipore)
- ChemiScreen
Qty/Pk:
200 units
Product Overview:
Human sst1
Background Information:
Somatostatin, a 14 or 28 amino acid peptide, inhibits hormone secretion from exocrine glands, and displays an antiproliferative activity on a number of cell types. A family of 5 Gi-coupled receptors mediates the effects of somatostatin. The classical activity of somatostatin of inhibiting growth hormone release from the pituitary appears to be mediated by sst1. In addition, sst1 functions as an inhibitory autoreceptor on somatostatin neurons in the hippocampus, hypothalamus, basal ganglia and retina (Thermos et al., 2006). The somatostatin receptors are targets for anticancer drugs and for treatment of hormonal dysfunction. Millipore's sst1 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 sst1. The membrane preparations exhibit a Kd of 1.4 nM for [125I]- somatostatin-14. With 0.5 nM [125I]- somatostatin-14, 5 μg/well sst1 Membrane Prep yields greater than 12 fold signal-to-background ratio.
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.
Species:
Human
Quality Assurance:
Table 1. Signal:background and specific binding values obtained in a competition binding assay with varying amounts of sst1 membrane prep.
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 8.4 pmol/mg
Kd: 1.4 nM
| 5 µg/well | |
|---|---|
| Signal:Background | 20.4 |
| Specific Binding (cpm) | 14,647 |
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 8.4 pmol/mg
Kd: 1.4 nM
Cell Type:
Chem-1
Brand Family:
Chemicon
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, 0.5% BSA. 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, 0.2% BSA, filtered and stored at 4°C
Radioligand: [125I]- somatostatin-14 (PerkinElmer NEX389)
Wash Buffer: 50 mM HEPES, pH 7.4, 500 mM NaCl, 0.1% BSA, filtered and stored at 4°C.
Binding buffer: 50 mM HEPES, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.2% BSA, filtered and stored at 4°C
Radioligand: [125I]- somatostatin-14 (PerkinElmer NEX389)
Wash Buffer: 50 mM HEPES, pH 7.4, 500 mM NaCl, 0.1% BSA, filtered and stored at 4°C.
Host Cells:
Chem-1
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 12-fold signal:background with [125I]- somatostatin-14 at 0.5 nM.
Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives.
Packaging method: Membrane protein was 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: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Protein Target:
sst1
Ligand Type:
peptide
Storage Conditions:
Store at –70°C. Product is stable for at least 6 months from the date of receipt when stored as directed. Do not freeze and thaw.
Target Sub-Family:
Somatostatin
Promotional Text:
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GPCR Class:
A
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiSCREEN™ Membrane Preparation Recombinant Human sst1 Somatostatin Receptor
Entrez Gene Number:
Cell Membrane Type:
GPCR