ChemiScreen™ Human Recombinant LPA₃ Lysophospholipid Receptor Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant LPA₃ Lysophospholipid Receptor Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human EDG7 cDNA encoding LPA₃

Lysophosphatidic acid (LPA) is a lysophospholipid produced by activated platelets that inhibits adenylate cyclase and stimulates DNA synthesis, changes in cell morphology, and increases in intracellular calcium in a variety of cultured mammalian cells. A family of three GPCRs, LPA₁, LPA₂ and LPA₃, mediates the biological effects of LPA (Contos et al., 2000). LPA₃ is expressed primarily in testes, prostate, heart and pancreas. Mice lacking LPA₃ have low expression of uterine COX-2 and display decreased uterine implantation and embryo spacing (Ye et al., 2005). Chemicon's cloned human LPA₃-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant LPA₃ 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 LPA₃ and its ligands.

Calcium flux assay, ligand binding assays

Human

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This gene encodes a member of the G protein-coupled receptor family, as well as the EDG family of proteins. This protein functions as a cellular receptor for lysophosphatidic acid and mediates lysophosphatidic acid-evoked calcium mobilization. This receptor couples predominantly to G(q/11) alpha proteins.

FUNCTION: SwissProt: Q9UBY5 # Receptor for lysophosphatidic acid (LPA), a mediator of diverse cellular activities. May play a role in the development of ovarian cancer. Seems to be coupled to the G(i)/G(o) and G(q) families of heteromeric G proteins.
SIZE: 353 amino acids; 40128 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Most abundantly expressed in prostate, testes, pancreas, and heart, with moderate levels in lung and ovary. No detectable expression in brain, placenta, liver, skeletal muscle, kidney, spleen, thymus, small intestine, colon, or peripheral blood leukocytes.
SIMILARITY: SwissProt: Q9UBY5 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by Oleoyl-L-α-lysophosphatidic acid: ~ 520nM

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.

LPA3 / EDG7

Q9UBY5

non-peptide

Lysophospholipid

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.

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A

2x10⁶ cells/vial

• EDG7
• LP-A3
• LPA-3
• RP4-678I3
• Edg-7
• HOFNH30
• LPA3
• LPAR3
• GPCR

GPCRs

ChemiScreen™ Human Recombinant LPA₃ Lysophospholipid Receptor Calcium-Optimized Stable Cell Line

NM_012152.1

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.