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HTS025M ChemiSCREEN™ Membrane Preparation Recombinant Human MCH1 Receptor

Human MCHR1 / GPR24 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Host Cell: Chem-1 Uni Prot Number: Q99705 Entrez Gene Number: NM_005297
HTS025M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1MCHR1 / GPR24
Description
Catalogue NumberHTS025M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human MCH1 Receptor
OverviewFull length human MCHR1 cDNA encoding MCH1
Background InformationMelanin-concentrating hormone (MCH) is a cyclic, 19 amino acid peptide produced in the lateral hypothalamus and other regions of the mammalian brain (Bittencourt et al., 1992). When administered intracerebrally or overexpressed transgenically in rodents, MCH stimulates food intake and weight gain (Ludwig et al., 2001). A receptor for MCH, previously identified as an orphan receptor SLC-1 and renamed MCH1, has been identified in humans and rodents. Consistent with a role in hypothalmic control of feeding behavior, MCH1 is expressed in in the ventromedial and dorsomedial nuclei of the hypothalamus (Chambers et al., 1999). Genetic ablation and pharmacological antagonism of MCH1 in rodents results in resistance to obesity caused by a high fat diet or leptin deficiency (Borowsky et al., 2002; Segal-Lieberman, et al., 2003; Shearman et al., 2003; Marsh et al., 2002; Chen et al., 2002). Thus, MCH1 is an attractive target for obesity. Millipore's MCH1membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level GPCR surface expression; thus, they are ideal HTS tools for screening for agonists and antagonists of MCH1. The membrane preparations exhibit a Kd of 0.41 nM for [125I]-MCH. With 0.4 nM for [125I]-MCH, 5 μg/well MCH1 Membrane Prep yields greater than 8 fold signal-to-background ratio.
References
Product Information
FormatMembranes
PresentationOne package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 8-fold signal:background with 125I-labeled MCH at 0.4 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.
Quality LevelMQ100
Applications
ApplicationHuman MCHR1 / GPR24 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications
  • Radioligand binding assay
  • GTPγS binding
Application NotesRadioligand binding assay and GTPγS binding
Biological Information
Entrez Gene Number
Gene Symbol
  • MCHR1
  • GPR24
  • SLC1
GPCR ClassA
Host CellsChem-1
Protein TargetMCHR1 / GPR24
Target Sub-FamilyMelanin-concentrating hormone
UniProt Number
Incubation ConditionsRECOMMENDED 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]-MCH (Perkin Elmer NEX373)

Wash Buffer: 50 mM Hepes, pH 7.4, 500mM NaCl , 0.1% BSA, filtered and stored at 4°C
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Table 1. Signal:background and specific binding values obtained in a competition binding assay with MCH1 membrane prep.
Product Usage Statements
5 µg/well
Signal:Background 13.9
Specific Binding (cpm) 7776

SPECIFICATIONS: 1 unit = 5 µg membrane preparation
Bmax: 1.9 pmol/mg
Kd: 0.41 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 ConditionsStore 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.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS025M 04053252676284

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