ChemiScreen™ Human recombinant TRH Thyrotropin-Releasing Hormone Receptor, Calcium-Optimized Stable Cell Line

ChemiScreen™ Human recombinant TRH Thyrotropin-Releasing Hormone Receptor, Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

TRANSFECTION: Full-length human TRHR cDNA encoding TRH

Thyrotropin-releasing hormone (TRH) is a tripeptide (L-pyroglutamyl-L-histidinyl-L-proline amide) that is released from the hypothalamus to stimulate release of thyroid-stimulating hormone from the thyrotropes of the anterior pituitary. In addition, TRH is synthesized in other regions of the CNS, where it appears to function as a neuromodulator to influence arousal, cognition, motor activity, depression, and food and water intake. TRH functions in the autonomic nervous system to modulate gastrointestinal motility and acid secretion (Nillni and Severino, 1999). The biological functions of TRH are mediated by binding to two Gq-coupled seven transmembrane receptors, TRH1 and TRH2 (Sun et al., 2003). TRH1 is present in humans, rodents, and other vertebrates, whereas TRH2 has been identified only in rodents. Synthetic TRH (protirelin) is used clinically to evaluate the functionality of the hypothalamic-pituitary-thyroid axis. Chemicon's cloned human TRH -expressing cell line is made in the Chem-1 host, which supports high levels of recombinant TRH 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 TRH and its ligands.

Calcium flux assay, ligand binding assays

Human

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FUNCTION: SwissProt: P34981 # Receptor for thyrotropin-releasing hormone. This receptor is mediated by G proteins which activate a phosphatidylinositol-calcium second messenger system.
SIZE: 398 amino acids; 45085 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: P34981 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by TRH: ~ 4.8 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.

TRH

Cell Line

P34981

peptide

TRH

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

• TRHR
• MGC141920
• TRH-R

GPCRs

ChemiScreen™ Human recombinant TRH Thyrotropin-Releasing Hormone Receptor, Calcium-Optimized Stable Cell Line

NM_003301.2

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

2 x 10⁶ cells/mL