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ChemiScreen™ Membrane Preparation Human Recombinant k (kappa)

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号:HTS095M

牌:Millipore 密理博

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Replacement Information

重要规格表

Species Host Cells Protein Target
Human Chem-1 Kappa / OP2 / KOP / KOR
Description
Catalogue Number HTS095M
Brand Family Chemicon®
Trade Name
  • ChemiScreen
  • Chemicon
Description ChemiScreen™ Membrane Preparation
Recombinant Human Kappa Opioid Receptor
Overview Human OPK1, κ
Background Information Opiates derived from the opium poppy, Papaver somniferum, have been used in for millenia for their anti-diarrheal, analgesic and euphoric properties. More recently, endogenous peptides, enkephalins, dynorphins, and endorphins, were found to bind to the same sites as opiate alkaloids. The receptors for the classical opioids are three related GPCRs, µ, κ, and δ, that activate Gi/o to reduce intracellular cAMP levels. Most clinically used opioids function by activation of the µ opioid receptor. Activation of the κ opioid receptor by selective agonists also produces analgesia, primarily mediated by spinal sites, but causes dysphoria and psychosis instead of euphoria. The κ receptor at central and peripheral sites is also largely responsible for the anti-diarrheal effects of opiates (Dhawan et al., 1996). Chemicon's 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 antagonists of κ interactions with diprenorphine. The membrane preparations exhibit a Kd of 2.3-3.7 nM for [3H]-diprenorphine. With 10 µg/well κ Membrane Prep and 2.5 nM [3H]-diprenorphine, a greater than 3-fold signal-to-background ratio was obtained.
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 3-fold signal:background with 3H-labeled diprenorphine at 2.5 nM.

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 packaging buffer, rapidly frozen, and stored at -80°C.
Applications
Application Human Kappa / OP2 / KOP / KOR 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
Gene Symbol
  • OPRK1
  • OPRK
  • KOR-1
  • KOR
GPCR Class A
Host Cells Chem-1
Protein Target Kappa / OP2 / KOP / KOR
Target Sub-Family Opioid
UniProt Number
UniProt Summary FUNCTION: SwissProt: P41145 # Inhibits neurotransmitter release by reducing calcium ion currents and increasing potassium ion conductance. Receptor for dynorphins. May play a role in arousal and regulation of autonomic and neuroendocrine functions.
SIZE: 380 amino acids; 42645 Da
SUBUNIT: Binds SLC9A3R1.
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P41145 ## Belongs to the G-protein coupled receptor 1 family.
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 Binding Buffer. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Binding Buffer. The plate is dried and counted.

Binding buffer: 50 mM Tris-HCl pH 7.4, filtered and stored at 4°C
Radioligand: [3H] diprenorphine (Perkin Elmer # NET1121)
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information

SPECIFICATIONS: Signal:background and specific binding values obtained in a competition binding assay with κ membrane prep:
Product Usage Statements
10 µg/well
Signal:Background 4.47
Specific Binding (cpm) 939


1 unit = 10 µg membrane preparation
Bmax: 21.73 pmol/mg
Kd: 3.0 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 Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.
Packaging Information
Material Size 200 units
Transport Information
Supplemental Information
Specifications

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