ChemiScreen™ Human Recombinant GPR68/OGR1 Proton-Sensing Receptor

ChemiScreen™ Human Recombinant GPR68/OGR1 Proton-Sensing Receptor

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human GPR68 cDNA encoding OGR1

GPR68, also known as OGR1 (ovarian cancer G-protein-coupled receptor 1), was initially thought to be a receptor for sphingosylphosphorylcholine, although these results are controversial. Subsequent studies indicated that GPR68/OGR1 functions as a Gq-coupled sensor for extracellular pH, with maximal signaling occurring at pH of 7 or below (Ludwig et al., 2003; Tomura et al., 2005). Expression of GPR68/OGR1 is upregulated during RANKL-induced osteoclast differentiation, and knockdown of GPR68/OGR1 expression inhibits this differentiation process (Yang et al., 2006). Chemicon's cloned human GPR68-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant GPR68 expression on the cell surface and contains high levels of the promiscuous G protein Gα15 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 GPR68 and its ligands.

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: Q15743 # High affinity receptor for sphingosylphosphorylcholine, coupled to G proteins that stimulate phosphoinositide hydrolysis.
SIZE: 365 amino acids; 41077 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Found in many tissues, including spleen, testis, peripheral blood leukocytes, brain, lung, and placenta. Not detectable in ovary.
SIMILARITY: SwissProt: Q15743 ## Belongs to the G-protein coupled receptor 1 family.

Human

EC50 for calcium mobilization by pH: 6.4

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.

GPR68/OGR1

Q15743

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 37°C 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.

SPC/LPC /proton-sensor

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A

2 x 10⁶ cells/vial

• GPR68
• MGC111379
• OGR1
• OGR-1
• GPR12A

GPCRs

ChemiScreen™ Human Recombinant GPR68/OGR1 Proton-Sensing Receptor

NM_003485.3

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.