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HTS040M ChemiSCREEN™ Membrane Preparation Recombinant Human NOP Opioid Receptor

Human NOP / ORL1 / OP4 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: P35377 Entrez Gene Number: X72304
HTS040M
200 units  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

SpeciesHost CellsProtein Target
Human Chem-1NOP / ORL1 / OP4
Description
Catalogue NumberHTS040M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
DescriptionChemiSCREEN™ Membrane Preparation Recombinant Human NOP Opioid Receptor
OverviewFull-length human OPRL1 cDNA encoding NOP
Background InformationThe NOP receptor (also known as ORL1) is related to the opioid receptor family of GPCRs but does not bind to classical opioids. An endogenous ligand for NOP has been characterized and termed orphanin FQ or nociceptin (OFQ/N), which in turn does not bind to other members of the opioid receptor family. NOP is expressed widely in the CNS, and binding of OFQ/N to NOP1 appears to function in nociception, locomotor activity, anxiety, reward, memory and tolerance to classical opioids (Mogil and Pasternak, 2001). Millipore's NOP 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 agonists and antagonists of NOP. The membrane preparations exhibit a Kd of 0.06 nM for [125II]-Nociceptin. With 0.1 nM [125I]-Nociceptin, 5μg/well NOP Membrane Prep typically yields greater than 10-fold signal-to-background ratio.
References
Product Information
FormatMembranes
PresentationLiquid 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 1 ml packaging buffer, rapidly frozen, and stored at -80°C
Quality LevelMQ100
Applications
ApplicationHuman NOP / ORL1 / OP4 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
  • OPRL1
  • OOR
  • ORL1
GPCR ClassA
Host CellsChem-1
Protein TargetNOP / ORL1 / OP4
Target Sub-FamilyOpioid
UniProt Number
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, an FC 96-well harvest plate (Millipore cat. # MAHF C1H) is coated with 0.33% polyethyleneimine for 30 min, then washed with 50 mM Tris-HCl, pH 7.4. 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 Tris-HCl, pH 7.4 filtered and stored at 4°C

Radioligand: [125I]-Nociceptin. (Perkin Elmer#:NEX-324 )

Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl , 0.1% BSA, filtered and stored at 4°C.

One package contains enough membranes for at least 200 assays (units), where a unit is the amount of membrane that will yield greater than 10-fold signal:background with 125I labeled Nociceptin
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 NOP Receptor membrane prep.
Product Usage Statements
5 µg/well
Signal:Background 23.8
Specific Binding (cpm) 6656


SPECIFICATIONS: 1 unit = 5 µg
Bmax for [125I]-Nociceptin binding: 1.1 pmol/mg protein
Kd for [125I]-Nociceptin binding: ~0.06 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 up to the expiration date indicated on the product label. Do not freeze and thaw.
Packaging Information
Material Size200 units
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
HTS040M 04053252271250

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