ChemiScreen™ DP Prostanoid Receptor Calcium-Optimized Cell Line

ChemiScreen™ DP Prostanoid Receptor Calcium-Optimized Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human PTGDR cDNA encoding DP

Prostanoids are a series of arachidonic acid metabolites produced by the action of cyclooxygenase and subsequently by isomerases and synthases. Cells rapidly secrete prostanoids after synthesis, whereupon the prostanoids bind to a family of 8 GPCRs to exert their biological effects (Narumiya and FitzGerald, 2001). The prostaglandin PGD2 is produced by mast cells upon activation by allergens, and is present at high levels in allergic diseases. PGD2 binds to two receptors, DP and CRTH2. DP activates Gs to increase cAMP levels, and lack of DP results in reduced allergic response in animal models of bronchial asthma (Matsuoka et al., 2000). Chemicon's cloned human DP-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant DP expression on the cell surface and contains high levels of the promiscuous G protein Gα15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between DP and its ligands.

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.

The protein encoded by this gene is a G-protein-coupled receptor. It has been shown to function as a prostanoid DP receptor. The activity of this receptor is mainly mediated by G-S proteins that stimulate adenylate cyclase resulting in an elevation of intracellular cAMP and Ca2+. Knockout studies in mice suggest that the ligand of this receptor, prostaglandin D2 (PGD2), functions as a mast cell-derived mediator to trigger asthmatic responses.

FUNCTION: SwissProt: Q13258 # Receptor for prostaglandin D2 (PGD2). The activity of this receptor is mainly mediated by G(s) proteins that stimulate adenylate cyclase, resulting in an elevation of intracellular cAMP. A mobilization of calcium is also observed, but without formation of inositol 1,4,5-trisphosphate.
SIZE: 359 amino acids; 40271 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Expressed in retina and small intestine.
DISEASE:SwissProt: Q13258 # Genetic variations in PTGDR are associated with susceptibility to asthma-related traits type 1 (ASRT1) [MIM:607277]. Asthma-related traits include clinical symptoms of asthma, such as coughing, wheezing and dyspnea.
SIMILARITY: SwissProt: Q13258 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS:
EC50 for calcium mobilization by PGD2: ~ 38 nM
IC50 for BW-A868C: 25-60 nM
Signal/noise ratio at agonist Emax: 468
Z' with PGD₂ at 2x EC50: 0.86

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.

DP

Cell Line

Q13258

non-peptide

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.

Prostanoid

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

A

• PTGDR
• DP
• MGC49004
• AS1
• ASRT1

GPCRs

ChemiScreen™ DP Prostanoid Receptor Calcium-Optimized Cell Line

NM_000953.2

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.