ChemiSCREEN™ Human MC₄ Melanocortin Receptor Membrane Preparation

ChemiSCREEN™ Human MC₄ Melanocortin Receptor Membrane Preparation

• ChemiScreen
• Chemicon (Millipore)

200 units

Full-length human MC4R cDNA encoding for MC4 gene

The melanocortins, α-, β- and γ-melanocyte-stimulating hormones (MSHs), are peptides derived from a precursor protein POMC. The MSH peptides bind to a family of five Gs-coupled seven transmembrane recptors (MCRs) and play important roles in energy balance, reproductive function, pigmentation and inflammation (Gantz and Fong, 2003). The activity of the melanocortins is modulated by endogenous antagonist proteins, agouti and agouti-related protein (AGRP). MC4R is expressed primarily in the CNS and appears to play a prominent role in energy homeostasis. The hypothalamus, which is a major central site for control of feeding behavior, prominently expresses MC4R. Targeted deletion of MC4R in mice and naturally occurring mutations in MC4R in humans result in obese phenotypes. In addition, blockade of MC4R function by antagonists and targeted deletion of the gene in mice reverses melanocortin agonist-induced inhibition of food intake and promotes weight gain in uremia and cancer-induced cachexia (Cheung et al., 2005; Markison et al., 2005). Chemicon's MC4 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 MC4 interactions with αMSH. The membrane preparations exhibit a Kd of 0.71-0.77 nM for [¹²⁵I]-NDP-αMSH. With 10 µg/well MC4 Membrane Prep and 0.5 nM [¹²⁵I]-NDP-αMSH, a greater than 15-fold signal-to-background ratio was obtained.

Radioligand binding assay and GTPγS binding.

Human

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.

FUNCTION: SwissProt: P32245 # Receptor specific to the heptapeptide core common to adrenocorticotropic hormone and alpha-, beta-, and gamma-MSH. This receptor is mediated by G proteins that stimulate adenylate cyclase.
SIZE: 332 amino acids; 36943 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Brain, placental, and gut tissues.
DISEASE:SwissProt: P32245 # Defects in MC4R are a cause of autosomal dominant obesity [MIM:601665].
SIMILARITY: SwissProt: P32245 ## Belongs to the G-protein coupled receptor 1 family.

Human

Signal:background and specific binding values obtained in a competition binding assay with varying amounts of MC4 membrane prep:

	10 µg/well
Signal:Background	26.7
Specific Binding (cpm)	10325

SPECIFICATIONS: 1 unit = 10 μg membrane preparation
Bmax: 5.5 pmol/mg
Kd: 0.74 nM

Chem-3

Chemicon

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, 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 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] NDP-αMSH (Perkin Elmer # NEX352)
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 an unit is the amount of membrane that will yield greater than 15-fold signal:background with ¹²⁵I-labeled NDP-αMSH at 0.5 nM.

Chem-3

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, rapidly frozen, and stored at -80°C.

MC4

P32245

peptide

Melanocortin

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

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A

2 mL, 1 mg/mL

• MC4R
• MC4-R
• MGC138197
• MGC126851

GPCR Membrane Preparations

ChemiSCREEN™ Human MC₄ Melanocortin Receptor Membrane Preparation

NM_005912.2

MC4R

GPCR