ChemiScreen™ GAL1 Calcium-Optimized FLIPR Cell Lines

ChemiScreen™ GAL1 Calcium-Optimized FLIPR Cell Lines

• ChemiScreen
• Chemicon (Millipore)

2 vials

Galanin is a 29-30 amino acid peptide originally purified from intestine but later found to be abundant in the CNS. A family of 3 GPCRs, GAL1, GAL2 and GAL3, bind to galanin and mediate its biological effects. GAL1 couples to Gi/o to decrease intracellular cAMP levels (Wang et al., 1998). Galanin and its receptors have been implicated in pain, cognition, seizure activity, depression and drug addiction (Hökfelt, 2005). In particular, GAL1 mediates the anticonvulsant activity of galanin; mice lacking GAL1 are affected by spontaneous seizures, and have increased susceptibility to seizures induced by lithium-pilocarpine treatment and electrical perforant path stimulation (McColl et al., 2006; Mazarati et al., 2004). Chemicon's cloned human GAL1-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant GAL1 expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between GAL1 and its ligands.

Calcium flux assay, ligand binding assays

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The neuropeptide galanin elicits a range of biological effects by interaction with specific G-protein-coupled receptors. Galanin receptors are seven-transmembrane proteins shown to activate a variety of intracellular second-messenger pathways. GALR1 inhibits adenylyl cyclase via a G protein of the Gi/Go family. GALR1 is widely expressed in the brain and spinal cord, as well as in peripheral sites such as the small intestine and heart.

FUNCTION: SwissProt: P47211 # Receptor for the hormone galanin. The activity of this receptor is mediated by G proteins that inhibit adenylate cyclase activity.
SIZE: 349 amino acids; 38897 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
PTM: Palmitoylated on at least one of the three cysteine residues present in the C-terminal part.
SIMILARITY: SwissProt: P47211 ## Belongs to the G-protein coupled receptor 1 family.

Human

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

Cells are frozen at 2 x 10e6 cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

GAL1

P47211

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 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.

Galanin

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A

1 mL per vial

• GALR1
• GALNR1
• GAL1-R
• GALNR

GPCRs

ChemiScreen™ GAL1 Calcium-Optimized FLIPR Cell Lines

NM_001480.2

2 x 10e6 cells/mL