Content Loading...
Content Loading...
Product Images
Figure 2. Competition binding for Ghrelin Receptor. 5µg/well or 10g/well Ghrelin Receptor Membrane Preparation and wild-type Chem-1 Membrane Prepar...
Figure 1. Saturation binding for Ghrelin Receptor. 5.0 µg/well Ghrelin Receptor Membrane Preparation was incubated with increasing amount of [1...
ChemiSCREEN™ Membrane Preparation Recombinant Human Gherlin Receptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human Gherlin Receptor
Trade Name:
Chemicon (Millipore)
Qty/Pk:
200 units
Product Overview:
Human GHSR cDNA encoding the ghrelin receptor isoform 1a
Background Information:
Ghrelin is a 28 amino acid peptide, containing a unique octanoyl moiety added post-translationally to Ser3, with diverse activities in the CNS, gastrointestinal tract, and cardiovascular system (Davenport et al., 2005; Leite-Moreira and Soares, 2007). In the CNS, ghrelin stimulates release of growth hormone from the anterior pituitary and increases appetite by binding to neurons within the arcuate nucleus. Circulating concentrations of ghrelin increase with preprandially and decrease post-prandially, and thus counterbalances the effects of leptin to coordinate energy balance, appetite and food intake. Ghrelin is also expressed in the cardiovascular system where it acts as a potent vasodilator; receptors are up-regulated in patients with atherosclerosis, suggesting that it plays a role in compensating for increased vasoconstriction (Kleinz et al., 2006). The effects of ghrelin are mediated by a Gq-coupled receptor, originally designated GHSR (growth hormone secretogogue receptor), and more recently termed the Ghrelin Receptor or GRLN. Two splice variants have been described; type 1a (GHS-R1a) is the functional receptor, whereas type 1b (GHS-R1b) encodes a truncated, inactive protein with only 5 transmembrane domains. As administration of agonists of the ghrelin receptor to rats leads to increased food intake and antagonists reduce food intake (Beck et al. 2004), antagonism and inverse agonism of the ghrelin receptor may reduce food intake in certain types of obesity, and agonists of the ghrelin receptor are potentially useful for treatment of anorexia and cachexia. Millipore's Ghrelin Receptor 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 ghrelin interactions and its ligands. The membrane preparations exhibit a Kd of 0.32 nM for [125I]-ghrelin. With 0.1 nM [125I]-ghrelin, 5 μg/well Ghrelin Receptor Membrane Prep typically 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 various amount of membrane prep.
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 4.2 pmol/mg
Kd: 0.32 nM
| 5 µg/well | 10 µg/well | |
|---|---|---|
| Signal:Background | 19.7 | 17.2 |
| Specific Binding (cpm) | 9228 | 9249 |
SPECIFICATIONS: 1 unit = 5 μg membrane preparation
Bmax: 4.2 pmol/mg
Kd: 0.32 nM
Brand Family:
Chemicon
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 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]-Ghrelin. (Perkin Elmer#:NEX-338 )
Wash Buffer: 50 mM Hepes, 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 12-fold signal:background with 125I labeled ghrelin.
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]-Ghrelin. (Perkin Elmer#:NEX-338 )
Wash Buffer: 50 mM Hepes, 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 12-fold signal:background with 125I labeled ghrelin.
Host Cells:
Chem-1
Presentation:
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.
Packaging method: Membrane protein was adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80°C.
Protein Target:
Ghrelin / GHSR-1a / growth hormone secretagogue receptor
Format:
Membranes
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:
Ghrelin
Promotional Text:
For special offers Click here http://www.millipore.com/drugdiscovery/dd3/gpcrtargetsolutions
GPCR Class:
A
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiSCREEN™ Membrane Preparation Recombinant Human Gherlin Receptor
Entrez Gene Number:
Cell Membrane Type:
GPCR