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HTS022M ChemiSCREEN™ MC3 Membrane Preparation

Human MC3 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.

ChemiSCREEN™ MC3 Membrane Preparation MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: P41968 Entrez Gene Number: NM_019888.2
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HTS022M
200 assays  
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Overview

Replacement Information

Key Specifications Table

SpeciesHost CellsProtein Target
Human Chem-1MC3
Description
Catalogue NumberHTS022M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ MC3 Membrane Preparation
OverviewHuman MC3
Background InformationThe melanocortins, the peptides alpha-, beta- and gamma-melanocyte-stimulating hormone (MSH), bind to a family of five Gs-coupled GPCR receptors (MCRs) and play important roles in energy balance, reproductive function, pigmentation and inflammation (Gantz and Fong, 2003). MC3R is expressed in the CNS, including the hypothalamus, and appears to play a role in energy homeostasis as indicated by the observation that MC3R-null mice have increased subcutaneous fat, a loss of lean body mass, and reduced energy expenditure (Butler et al., 2000; Chen et al., 2000). Macrophages also express MC3R, which together with MC1R mediates the anti-inflammatory activity of melanocortins in animal models of peritonitis and myocardial infarct (Getting et al., 2003; 2004). Chemicon's MC3R 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 MC3R interactions with melanocortins. The membrane preparations exhibit a Kd of 0.4 nM for [125I]-MSH. With 10.0 mg/well MC3R Membrane Prep and 0.25 nM [125I]-MSH, a greater than 7-fold signal-to-background ratio is obtained.
References
Product Information
FormatMembranes
PresentationLiquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA no preservatives.Packaging method: Membranes protein were adjusted to 0.5 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C.
Quality LevelMQ100
Applications
ApplicationHuman MC3 GPCR membrane preparation for Radioligand binding Assays & GTPgammaS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay, and GTPgammaS binding.
Biological Information
Concentration0.5 mg/mL
Entrez Gene Number
Entrez Gene SummaryThis gene encodes a G-protein-coupled receptor for melanocyte-stimulating hormone and adrenocorticotropic hormone that is expressed in tissues other than the adrenal cortex and melanocytes. This gene maps to the same region as the locus for benign neonatal epilepsy. Mice deficient for this gene have increased fat mass despite decreased food intake, suggesting a role for this gene product in the regulation of energy homeostasis. Mutations in this gene are associated with a susceptibility to obesity in humans.
Gene Symbol
  • MC3R
  • MC3
  • MC3-R
GPCR ClassA
Host CellsChem-1
Protein TargetMC3
Target Sub-FamilyMelanocortin
UniProt Number
UniProt SummaryFUNCTION: SwissProt: P41968 # Receptor for MSH (alpha, beta and gamma) and ACTH. This receptor is mediated by G proteins which activate adenylate cyclase.
SIZE: 360 amino acids; 40073 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Brain, placental, and gut tissues.
SIMILARITY: SwissProt: P41968 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsMembranes 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 HEPES, pH 7.4, 0.5% BSA. Binding reaction is transferred to the filter plate, and washed 9 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.Binding buffer: 25 mM Hepes, pH 7.0, 1 mM MgCl2, 1.5 mM CaCl2 2H2O, 100mM NaCl, 0.2% BSA, Protease Inhibitor, filter and stored at 4 °C. Radioligand: [125I] MSH (Perkin Elmer# NEX352)

Wash Buffer: 25 mM Hepes, pH 7.0, 1 mM MgCl2, 1.5 mM CaCl2 2H2O, 100mM NaCl, 0.2% BSA, filtered and stored at 4°C. One vial 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 MSH at 0.25 nM
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Signal:background and specific binding values obtained in a competition binding assay with MC3 membrane prep and unlabeled NDP-MSH.
Product Usage Statements
10 µg/well
Signal:Background 7.7
Specific Binding (cpm) 5073


SPECIFICATIONS:
1 unit = 10 μg
Bmax: 0.77 pmol/mg
Kd: 1.0 nM 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.
Storage and Shipping Information
Storage ConditionsMaintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size200 assays
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalog Number GTIN
HTS022M 04053252511783

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