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HTS071M ChemiSCREEN™ Membrane Preparation Recombinant Human ChemR23 Chemoattractant Receptor

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HTS071M
200 units  
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Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1ChemR23 / CMKLR1
Description
Catalogue NumberHTS071M
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human ChemR23 Chemoattractant Receptor
OverviewFull-length human ChemR23
Background InformationChemR23 was discovered as an orphan receptor related to the chemoattractant receptors C3a, C5a and FPR1, and expressed on dendritic cells and macrophages (Samson et al., 1998). A ligand for ChemR23 was characterized as chemerin, a 15 kD proteolytically processed protein found in inflammatory sites; a 9 amino acid peptide from the C-terminus of chemerin is sufficient to activate ChemR23 (Wittamer et al., 2003, 2004). Chemerin expressed in lymphoid and microvascular endothelium mediates migration of ChemR23-expressing dendritic cells to lymphoid organs and vasculature at sites of inflammation (Vermi et al., 2005). In addition, a bioactive lipid, resolvin E1, was found to functionally interact with ChemR23 to reduce inflammation (Arita et al., 2005). Millipore's ChemR23 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 ChemR23 interactions with its ligands. The membrane preparations exhibit EC50s of 0.37 nM for chemerin in a GTPγS binding assay.
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 1 mg/ml in 1 ml packaging buffer, rapidly frozen, and stored at -80°C
Quality LevelMQ100
Applications
ApplicationHuman ChemR23 / CMKLR1 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
  • CMKLR1
  • ChemR23
  • DEZ
  • MGC126106
  • MGC126105
  • CHEMR23
GPCR ClassA
Host CellsChem-1
Protein TargetChemR23 / CMKLR1
Target Sub-FamilyChemoattractant
UniProt Number
UniProt SummaryFUNCTION: SwissProt: Q99788 # Orphan receptor. Could be a chemotactic peptide receptor. May have a function in bone metabolism. Acts as a coreceptor for several SIV strains (SIVMAC316, SIVMAC239, SIVMACL7E-FR and SIVSM62A), as well as a primary HIV-1 strain (92UG024-2).
SIZE: 373 amino acids; 42322 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Prominently expressed in developing osseous and cartilaginous tissue. Also found in adult parathyroid glands.
DEVELOPMENTAL STAGE: Expressed during bone development.
SIMILARITY: SwissProt: Q99788 ## Belongs to the G-protein coupled receptor 1 family.
Incubation ConditionsASSAY CONDITIONS: Membranes are permeabilized by addition of saponin to an equal concentration by mass, then mixed with [35S]-GTPγS (final concentration of 0.3 nM) in 20 mM HEPES, pH 7.4/100 mM NaCl/10 mM MgCl2/0.5 μM GDP in a nonbinding 96-well plate. Unlabeled chemerin was added to the final concentration indicated in Figure 1 (final volume 100 μL), and incubated for 30 min at 30°C. The binding reaction is transferred to a GF/B filter plate (Millipore MAHF B1H) previously prewetted with water. The plate is washed 3 times (1 mL per well per wash) with cold 10 mM sodium phosphate, pH 7.4, then dried and counted.

One vial contains enough membranes for at least 200 assays (units), where one unit is the amount of membrane that will yield greater than 1000 cpm specific chemerin-stimulated [35S]-GTPγS binding.

The ChemR23 membrane preparation is expected to be functional in a radioligand binding assay; however, the end user will need to determine the optimal radiolabeled ligand for use with this product.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceSPECIFICATIONS: 1 unit = 5 μg
EC50 in GTPγS binding assay by Chemerin: ~ 0.37 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 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS071M 04053252468810

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Categories

Life Science Research > Drug Discovery and Development > GPCR Cell Lines & Assays > GPCR Membrane Preparations
Life Science Research > Proteins and Enzymes > GPCR Membrane Preparations