ChemiScreen™ Membrane Prep Hu Recombinant MCH1

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ChemiScreen™ Membrane Prep Hu Recombinant MCH1

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货号：HTS025M

品牌：Millipore 密理博

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Replacement Information

重要规格表

Species	Host Cells	Protein Target
Human	Chem-1	MCHR1 / GPR24

Description
Catalogue Number	HTS025M
Brand Family	Chemicon®
Trade Name	ChemiScreen Chemicon
Description	ChemiSCREEN™ Membrane Preparation Recombinant Human MCH₁ Receptor
Overview	Full length human MCHR1 cDNA encoding MCH₁
Background Information	Melanin-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
Format	Membranes
Presentation	One 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.

Product Information

Format

Membranes

Presentation

One 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.

Applications
Application	Human MCHR1 / GPR24 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.
Key Applications	Radioligand binding assay GTPγS binding
Application Notes	Radioligand binding assay and GTPγS binding

Biological Information
Entrez Gene Number	NM_005297
Gene Symbol	MCHR1 GPR24 SLC1
GPCR Class	A
Host Cells	Chem-1
Protein Target	MCHR1 / GPR24
Target Sub-Family	Melanin-concentrating hormone
UniProt Number	Q99705
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]-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

₁ Table 1. Signal:background and specific binding values obtained in a competition binding assay with MCH 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 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.