ChemiScreen™ Human recombinant SST₄ Somatostatin Receptor, Calcium-Optimized Stable Cell Line

ChemiScreen™ Human recombinant SST₄ Somatostatin Receptor, Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

TRANSFECTION: Full-length human SSTR₄ cDNA encoding SST₄.

Somatostatin is a 14 or 28 amino acid regulatory peptide that inhibits hormone secretion from the pituitary, pancreas, and other endocrine sites. A family of 6 GPCRs, sst₁, sst_2A, sst_2B, sst₃, sst₄ and sst₅, mediate the biological activity of somatostatins. The somatostatin receptors couple to Gi to inhibit cAMP production, and also increase MAP kinase signalling. Several tumors have been shown to overexpress somatostatin receptors, and binding of somatostatin to these tumor cells stimulates or inhibits proliferation, depending on the receptor subtypes expressed (Olias et al., 2004). Somatostatin has been implicated in seizure susceptibility in animal models, and activation of sst₄ with selective agonists increases seizure activity (Moneta et al., 2002). Chemicon's cloned human SST₄-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant SST₄ 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 SST₄ and its ligands.

Calcium flux assay, ligand binding assays

Human

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Somatostatin acts at many sites to inhibit the release of many hormones and other secretory proteins. The biologic effects of somatostatin are probably mediated by a family of G protein-coupled receptors that are expressed in a tissue-specific manner. SSTR4 is a member of the superfamily of receptors having seven transmembrane segments and is expressed in highest levels in fetal and adult brain and lung.

FUNCTION: SwissProt: P31391 # Receptor for somatostatin-14. The activity of this receptor is mediated by G proteins which inhibits adenylyl cyclase. It is functionally coupled not only to inhibition of adenylate cyclase, but also to activation of both arachidonate release and mitogen-activated protein (MAP) kinase cascade. Mediates antiproliferative action of somatostatin in tumor cells.
SIZE: 388 amino acids; 41895 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Specifically expressed in fetal and adult brain, lung tissue, stomach, and in lesser quantities in the kidney, pituitary and adrenals.
SIMILARITY: SwissProt: P31391 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by Somatostatin: ~ 0.4 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.

sst4

Cell Line

P31391

peptide

Somatostatin

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

• SSTR4
• SS4R

GPCRs

ChemiScreen™ Human recombinant SST₄ Somatostatin Receptor, Calcium-Optimized Stable Cell Line

NM_001052.2

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