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Figure 1. Saturation binding for MC5. 5 µg/well MC5 Membrane Preparation was incubated with increasing amount of [125
Figure 2. Competition binding for MC5. MC5 Membrane Preparation and Wild-Type Chem-1 membrane preparation (WT; Chemicon Catal...
ChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor
Description:
ChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor
Trade Name:
Chemicon (Millipore)
Qty/Pk:
200 units
Product Overview:
Full-length human MC5R cDNA encoding MC5
Background Information:
The melanocortin system consists of five seven-transmembrane spanning G-protein coupled receptors (MC1-MC5). Among the five members of the melanocortin receptor family, MC2 and MC5 are expressed in peripheral tissues. The MC2 receptor (ACTH receptor) is almost exclusively expressed in the adrenal cortex whereas MC5 is expressed in several organs including the adrenal cortex. Studies have shown that targeted disruption of the MC5R gene produced mice with a severe defect in water repulsion and thermoregulation caused by decreased production of sebaceous lipids (Chen et al., 1997). Data show a requirement for the MC5 in multiple exocrine glands for the production of numerous products, indicative of a coordinated system for regulation of exocrine gland function by melanocortin peptides. (Entwistle et al.,1990). Millipore's MC5 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 MC5. The membrane preparations exhibit a Kd of 1.3nM for [125I]-[Nle4, D-Phe7]-α-MSH (NDP-αMSH). With 0.3 nM [125I]-NDP-αMSH, 10µg/well MC5 Membrane Prep typically yields greater than 50-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.
UniProt Summary:
FUNCTION: SwissProt: P33032 # Receptor for MSH (alpha, beta and gamma) and ACTH. The activity of this receptor is mediated by G proteins which activate adenylate cyclase. This receptor is a possible mediator of the immunomodulation properties of melanocortins.
SIZE: 325 amino acids; 36601 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in the brain but not in the melanoma cells.
SIMILARITY: SwissProt: P33032 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 325 amino acids; 36601 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in the brain but not in the melanoma cells.
SIMILARITY: SwissProt: P33032 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Quality Assurance:
Table 1. Signal:background and specific binding values obtained in a competition binding assay with MC5 Receptor membrane prep.
SPECIFICATIONS: 1 unit = 10 µg
Bmax for [125I]-NDP-αMSH binding: 4.3 pmol/mg protein
Kd for [125I]-NDP-αMSH binding: ~1.3 nM
| 10 µg/well | |
|---|---|
| Signal:Background | 147.3 |
| Specific Binding (cpm) | 15208.7 |
SPECIFICATIONS: 1 unit = 10 µg
Bmax for [125I]-NDP-αMSH binding: 4.3 pmol/mg protein
Kd for [125I]-NDP-αMSH binding: ~1.3 nM
Cell Type:
Chem-1
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, 500mM NaCl. 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]-NDP-αMSH (Perkin Elmer#: NEX-352)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 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 50-fold signal:background with 125I labeled NDP-αMSH at 0.3 nM
Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl2, 1 mM CaCl2, filtered and stored at 4°C.
Radioligand: [125I]-NDP-αMSH (Perkin Elmer#: NEX-352)
Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl, 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 50-fold signal:background with 125I labeled NDP-αMSH at 0.3 nM
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: Membranes protein were adjusted to the indicated concentration in packaging buffer, and dispensed at 1 mL/vial. Vials were rapidly frozen, and stored at -80°C
Protein Target:
MC5
UniProt Number:
Ligand Type:
peptide
Storage Conditions:
Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Target Sub-Family:
Melanocortin
Promotional Text:
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GPCR Class:
A
Gene Symbol:
- MC5R
- MC5-R
- MC-2
Protein or Enzyme Type:
GPCR Membrane Preparations
Product Name:
ChemiSCREEN™ Membrane Preparation Recombinant Human MC5 Melanocortin Receptor
Entrez Gene Number:
Cell Membrane Type:
GPCR