ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE HUMAN RECOMBINANT OT OXYTOCIN RECEPTOR

ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE HUMAN RECOMBINANT OT OXYTOCIN RECEPTOR

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human OXTR cDNA encoding OT

Oxytocin is a cyclic 9 amino acid peptide that differs from vasopressin in 2 amino acids. Despite the close similarities in sequence, oxytocin and vasopressin have different biological activities and bind to distinct G protein-coupled receptors. The oxytocin receptor, OT, couples primarily to Gq/11 to mobilize intracellular calcium. In female reproduction, oxytocin promotes uterine contraction and lactation; oxytocin is the most commonly used drug for induction of labor, whereas an oxytocin antagonist, atosiban, is under investigation to suppress preterm labor. Oxytocin/OT interaction in the CNS also plays an important role in stress, male and female sexual response, and sociality (Gimpl and Fahrenholz, 2001). Chemicon's cloned human OT-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant OT 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 OT and its ligands.

Calcium flux assay, ligand binding assays

Human

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The protein encoded by this gene belongs to the G-protein coupled receptor family and acts as a receptor for oxytocin. Its activity is mediated by G proteins which activate a phosphatidylinositol-calcium second messenger system. The oxytocin-oxytocin receptor system plays an important role in the uterus during parturition.

FUNCTION: SwissProt: P30559 # Receptor for oxytocin. The activity of this receptor is mediated by G proteins which activate a phosphatidylinositol- calcium second messenger system.
SIZE: 389 amino acids; 42772 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P30559 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by oxytocin: ~ 1.2 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.

OT

P30559

peptide

Oxytocin

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 Ca²⁺ and Mg²⁺-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

1mL per vial

• OXTR
• OT-R

GPCRs

ChemiScreen™ CALCIUM-OPTIMIZED STABLE CELL LINE HUMAN RECOMBINANT OT OXYTOCIN RECEPTOR

NM_000916.3

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

2 x 10⁶ cells/mL