ChemiScreen™ Calcium-Optimized Stable Cell Line
Human Recombinant CGRP₁ Calcitonin Receptor-Like Receptor

ChemiScreen™ Calcium-Optimized Stable Cell Line
Human Recombinant CGRP₁ Calcitonin Receptor-Like Receptor

ChemiScreen

Full-length human CALCRL cDNA, encoding CGRP1, and RAMP1 cDNA.

The calcitonin family of peptides, which includes calcitonin, two calcitonin-related gene peptides (CGRPs), adrenomedullin, and amylin, share a disulfide-bonded loop, an amphipathic a-helix, and an amidated C-terminus. The peptides influence a variety of physiological effects, including vascular tone, food intake and bone metabolism. The biological effects of the peptides are mediated by two class B GPCRs, CT receptor (calcitonin receptor) and CL receptor (calcitonin receptor-like receptor) (Poyner et al., 2002). The pharmacology of the CT and CL receptors is strongly influenced by their association with a family of three receptor activity modifying proteins (RAMPs), which are 14-17 kD single pass transmembrane proteins. CGRP (calcitonin gene-related peptide) induces vasodilation, and its activity is primarily mediated by CGRP₁, composed of CL in complex with RAMP1 (Hasbak et al., 2003). As a consequence of its role as a vasodilator, CGRP acting via CGRP₁ appears to play an important role in hypertension (Deng and Li, 2005). Chemicon's cloned human CGRP₁-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant CGRP₁ 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 CGRP₁ and its ligands.

Calcium flux assay, ligand binding assays

Human

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.

1 mL per 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)/0.5 mg/mL Hygromycin/100 U/mL each penicillin and streptomycin.

Chem-1

• CALCRL
• CGRPR1-PEN
• CGRPR1
• CGRPR
• CRLR

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.

EC50 for calcium mobilization by ~CGRP: ~ 26.5 nM
EC50 for calcium mobilization by Adrenomedullin: ~ 310 nM

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 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 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.