Accéder au contenu
MilliporeSigma
Toutes les photos(2)

Principaux documents

HTS040M

Sigma-Aldrich

ChemiSCREEN Membrane Preparation Recombinant Human NOP Opioid Receptor

Human NOP / ORL1 / OP4 GPCR membrane preparation for Radioligand binding Assays & GTPγS binding.

Se connecterpour consulter vos tarifs contractuels et ceux de votre entreprise/organisme


About This Item

Code UNSPSC :
41106514
eCl@ss :
32161000
Nomenclature NACRES :
NA.84

Source biologique

human

Niveau de qualité

Produit recombinant

expressed in Chem-1 cells

Fabricant/nom de marque

ChemiScreen
Chemicon®

Technique(s)

ligand binding assay: suitable (GTPγS)
radioligand binding assay (RLBA): suitable

Numéro d'accès UniProt

Conditions d'expédition

dry ice

Description générale

Full-length human OPRL1 cDNA encoding NOP
The 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.

Application

Radioligand binding assay and GTPγS binding

Actions biochimiques/physiologiques

GPCR Class: A
Protein Target: NOP / ORL1 / OP4
Target Sub-Family: Opioid

Qualité

Table 1. Signal:background and specific binding values obtained in a competition binding assay with NOP Receptor membrane prep.
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

Caractéristiques

Inucbation 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 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

Forme physique

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 1 ml packaging buffer, rapidly frozen, and stored at -80°C

Stockage et stabilité

Maintain frozen at –70°C up to the expiration date indicated on the product label. Do not freeze and thaw.

Informations légales

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Clause de non-responsabilité

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.

Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 2

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable


Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

Déjà en possession de ce produit ?

Retrouvez la documentation relative aux produits que vous avez récemment achetés dans la Bibliothèque de documents.

Consulter la Bibliothèque de documents

Notre équipe de scientifiques dispose d'une expérience dans tous les secteurs de la recherche, notamment en sciences de la vie, science des matériaux, synthèse chimique, chromatographie, analyse et dans de nombreux autres domaines..

Contacter notre Service technique