ChemiSCREEN™ CRF2 Calcium-Optimized FLIPR Cell Line

ChemiSCREEN™ CRF2 Calcium-Optimized FLIPR Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human CRF2 cDNA

The corticotropin-releasing factor receptors, CRF1 and CRF2, are Gs-coupled GPCRs expressed in the brain, blood vessels and intestine that bind to several neuropeptides, including corticotropin-releasing factor (CRF) and urocortin, and the amphibian peptide sauvagine (Lovenberg et al., 1995; Bale and Vale, 2004). In addition, two peptides, urocortin II (Ucn II) and urocortin III (Ucn III), bind selectively and with high affinity to CRF2 (Lewis et al., 2001). The CRF peptides and their receptors play important roles in stress mediated by the hypothalamic-pituitary-adrenal axis in animal models, and possibly in depression and anxiety in humans, although the contributions of CRF1 and CRF2 appear to be distinct (Bale and Vale, 2004; Risbrough et al., 2004) In addition, CRF1 and CRF2 differentially alter feeding behavior, gastric motility and vascular tone (Zorrilla et al., 2004; Martinez et al., 2004; Wiley and Davenport, 2004). Chemicon's cloned human CRF2-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CRF2 expression on the cell surface and contains high levels of the promiscuous G protein Ga15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CRF2 and its ligands.

Calcium flux assay, ligand binding assays

Human

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FUNCTION: SwissProt: Q13324 # This is a receptor for corticotropin releasing factor. Shows high-affinity CRF binding. Also binds to urocortin I, II and III. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase.
SIZE: 411 amino acids; 47688 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
SIMILARITY: SwissProt: Q13324 ## Belongs to the G-protein coupled receptor 2 family.

Human

Chemicon

CRF2

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.

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 37°C 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 growth media, and replace media after 8-24 h. Cells are typically passaged 1:10 every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.

CRF Receptor

B

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• CRHR2
• CRH2R
• CRF2
• CRFR2
• CRF2R

GPCRs

ChemiSCREEN™ CRF2 Calcium-Optimized FLIPR Cell Line

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 10e6 cells/mL

SPECIFICATIONS: EC50 for calcium mobilization by Sauvagine: ~ 50 nM

Chem-1

GPCR Cell Lines

Chem-1

Cell Line

Q13324

2x10⁶ cells/vial

NM_001883.3