HTS170M
ChemiSCREEN Membrane Preparation Recombinant Human sst1 Somatostatin Receptor
Human sst1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
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About This Item
UNSPSC-Code:
41106514
eCl@ss:
32161000
NACRES:
NA.84
Biologische Quelle
human
Qualitätsniveau
Rekombinant
expressed in Chem-1 cells
Hersteller/Markenname
ChemiScreen
Chemicon®
Methode(n)
ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable
NCBI-Hinterlegungsnummer
UniProt-Hinterlegungsnummer
Versandbedingung
dry ice
Angaben zum Gen
human ... SSTR1(6751)
Allgemeine Beschreibung
Human sst1
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.
Anwendung
Human sst1 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Radioligand binding assay and GTPγS binding
Biochem./physiol. Wirkung
GPCR Class: A
Protein Target: sst1
Target Sub-Family: Somatostatin
Qualität
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
Spezifikationen
Inucbation 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.
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.
Physikalische Form
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.
Lagerung und Haltbarkeit
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.
Rechtliche Hinweise
CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany
Haftungsausschluss
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.
Lagerklassenschlüssel
12 - Non Combustible Liquids
WGK
WGK 2
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
Analysenzertifikate (COA)
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