ChemiScreen™ Kappa Calcium-Optimized FLIPR Cell Lines

ChemiScreen™ Kappa Calcium-Optimized FLIPR Cell Lines

• ChemiScreen
• Chemicon (Millipore)

2 vials

TRANSFECTION: Full-length human OPRK1 cDNA encoding the kappa opioid receptor

Opiates derived from the opium poppy, Papaver somniferum, have been used 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, kappa, and delta, that activate Gi/o to reduce intracellular cAMP levels. Most clinically used opioids function by activation of the μ opioid receptor. Activation of the kappa opioid receptor by selective agonists also produces analgesia, primarily mediated by spinal sites, but causes dysphoria and psychosis instead of euphoria. The kappa receptor at central and peripheral sites is also largely responsible for the anti-diarrheal effects of opiates (Dhawan et al., 1996). Chemicon's cloned human kappa-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant kappa expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists of the kappa receptor.

Calcium flux assay, ligand binding assays

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.

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.

Human

SPECIFICATIONS:
EC50 for calcium mobilization by U-50488: ~ 76.4 nM

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10⁶ cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

Kappa / OP2 / KOP / KOR

Cell Line

P41145

peptide

Opioid

Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2.. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca++ and Mg++-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.

For special offers Click here http://www.millipore.com/drugdiscovery/dd3/gpcrtargetsolutions

A

1 mL per vial

• OPRK1
• OPRK
• KOR-1
• KOR

GPCRs

ChemiScreen™ Kappa Calcium-Optimized FLIPR Cell Lines

NM_000912.3

GROWTH MEDIA: DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin

2 x 10⁶ cells/mL