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Figure 1. Calcium flux in CRF1-expressing Chem-1 cell line induced by Sauvagine. CRF1-expressing Chem-1 cells were loaded with ...
Figure 2. Assay for antagonist activity at CRF1 by calcium flux assay. CRF1-expressing Chem-1 cells were loaded with Fluo-4 an ...
ChemiSCREEN™ CRF1 Calcium-Optimized FLIPR Cell Line
Description:
ChemiSCREEN™ CRF1 Calcium-Optimized FLIPR Cell Line
Trade Name:
ChemiScreen
Product Overview:
Full-length human CRF1 cDNA
Background Information:
The CRF1 receptor is a Gs-coupled GPCR expressed in the brain and pituitary gland that binds to several neuropeptides, including corticotropin-releasing factor (CRF) and urocortin, and the amphibian peptide sauvagine (Chen et al., 1993; Dautzenberg and Hauger, 2002; Bale and Vale, 2004). CRF plays a predominant role in stress response mediated by the hypothalamic-pituitary-adrenal axis, and alterations in CRF and its receptors CRF1 and CRF2 appear to be linked to depression and anxiety (Holsboer, 1999; Bale and Vale, 2004). A number of small molecule antagonists of the CRF1 receptor have been characterized, including R121919, SC241, NBI27914, antalarmin, DMP-696, and CP 154,526. When delivered in animal models of psychiatric disorders, these antagonists display effectiveness in reducing stress-related behaviors (Kehne and De Lombaert, 2002). Chemicon's cloned human CRF1-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CXCR2 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 CRF1 and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays
Species Reactivity:
Human
Usage Statement:
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.
Cell Line Type:
GPCR Cell Lines
Host Cells:
Chem-1, an adherent cell line expressing the promiscuous G protein, Gα15.
Packaging:
2x10(e6) cells/vial
Materials Required but Not Delivered:
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
Cell Type:
Chem-1
Gene Symbol:
- CRHR1
- CRF1
- CRFR
- CRHR1f
- CRHR
- CRFR1
- CRF-R
- CRH-R1h
Format:
Cell Line
Presentation:
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.
Quality Assurance:
SPECIFICATIONS:
EC50 for calcium mobilization by Sauvagine: ~ 40 nM
IC50 for NBI27914: ~1.3 µM
IC50 for astressin: ~11 nM
Signal:noise at Sauvagine Emax: 340
EC50 for calcium mobilization by Sauvagine: ~ 40 nM
IC50 for NBI27914: ~1.3 µM
IC50 for astressin: ~11 nM
Signal:noise at Sauvagine Emax: 340
Protein or Enzyme Type:
GPCRs
Species:
Human
Storage Conditions:
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 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.