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Calcium flux in FP-expressing Chem-1 cell line induced by PGF2α. FP-expressing Chem-1 cells and Wild-Type Chem-1 cells (Chemicon catalog # HTSCHEM-1)...
ChemiScreen™ FP Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ FP Calcium-Optimized FLIPR Cell Lines
Trade Name:
- ChemiScreen
- Chemicon (Millipore)
Qty/Pk:
2 vials
Product Overview:
TRANSFECTION: Full-length human PTGFR cDNA encoding FP
Background Information:
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 PGF2alpha binds specifically to the FP receptor, which couples to Gq/11 to mobilize intracellular calcium. Binding of PGF2alpha to FP receptors in the corpus luteum is required for luteolysis and subsequent parturition in mice (Sugimoto et al., 1998). PGF2alpha also decreases intraocular pressure by an FP-dependent mechanism, and an PGF2--analog--latanoprost, is used clinically in the treatment of glaucoma (Crowston et al., 2004). FP also contributes to tachycardia induced by inflammatory stimuli (Takayama et al., 2005). Chemicon's cloned human FP-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant FP expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to enhance coupling of the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between FP and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays.
Species Reactivity:
Human
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.
Entrez Gene Summary:
The protein encoded by this gene is member of the G-protein coupled receptor family. This protein is a receptor for prostaglandin F2-alpha (PGF2-alpha), which is known to be a potent luteolytic agent, and may also be involved in modulating intraocular pressure and smooth muscle contraction in uterus. Knockout studies in mice suggest that the interaction of PGF2-alpha with this receptor may initiate parturition in ovarian luteal cells and thus induce luteolysis. Two transcript variants encoding different isoforms have been found for this gene.
UniProt Summary:
FUNCTION: SwissProt: P43088 # Receptor for prostaglandin F2-alpha (PGF2-alpha). The activity of this receptor is mediated by G proteins which activate a phosphatidylinositol-calcium second messenger system. Initiates luteolysis in the corpus luteum (By similarity).
SIZE: 359 amino acids; 40055 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P43088 ## Belongs to the G-protein coupled receptor 1 family.
SIZE: 359 amino acids; 40055 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P43088 ## Belongs to the G-protein coupled receptor 1 family.
Species:
Human
Brand Family:
Chemicon
Protein Target:
FP
Presentation:
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.
Ligand Type:
non-peptide
Storage Conditions:
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.
Target Sub-Family:
Prostanoid
GPCR Class:
A
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Gene Symbol:
- PTGFR
- MGC46203
- FP
- MGC120498
Protein or Enzyme Type:
GPCRs
Product Name:
ChemiScreen™ FP Calcium-Optimized FLIPR Cell Lines
Alternate Names:
FP PROSTANOID RECEPTOR
Materials Required but Not Delivered:
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.
Concentration:
2 x 106 cells/mL
Quality Assurance:
SPECIFICATIONS:
EC50 for calcium mobilization by PGF2alpha: ~ 45 nM
Signal/noise ratio at agonist Emax: 270
EC50 for calcium mobilization by PGF2alpha: ~ 45 nM
Signal/noise ratio at agonist Emax: 270
Cell Type:
Chem-1
Cell Line Type:
GPCR Cell Lines
Host Cells:
Chem-1
Format:
Cell Line
UniProt Number:
Packaging:
1 mL per vial
Entrez Gene Number: