ChemiScreen™ Human Recombinant Secretin Receptor Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant Secretin Receptor Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

Full-length human SCTR cDNA encoding the Secretin Receptor.

Secretin, a member of the secretin-glucagon peptide hormone family, is a 27 amino acid peptide that was originally isolated from the duodenum. In the duodenum, secretin is released to stimulate the release of digestive juices in the pancreas (Bayliss et al, 1902). The receptor for secretin is a class 2 (or class B) G protein coupled receptor that signals through G_s to stimulate cAMP production (Dong et al, 2002). Along with its traditional role in the pancreas, studies in secretin-deficient mice have shown miscommunication between the CA3 Schaffer collateral and CA1 pyramidal neurons, causing a deficiency in synaptic transmission (Nishijima et al, 2006). This miscommunication of CA1 dendrites is found in Autism, Rett Syndrome, and most forms of mental retardation, suggesting secretin could be a potential target for treatment of these disorders. Chemicon's cloned human Secretin Receptor-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant Secretin Receptor 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 the Secretin Receptor 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.

GPCR Cell Lines

Chem-1, an adherent cell line expressing the promiscuous G protein, Gα15.

2 x 10⁶ cells/vial

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.

Chem-1

• SCTR
• SR
• SCT-R

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.

SPECIFICATIONS: EC50 for calcium mobilization by Secretin: ~ 19.5nM

GPCRs

Human

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.