ChemiScreen™ Human Recombinant CB₂ Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant CB₂ Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

Cannabinoid compounds include exogenous drugs such as δ9-THC, the main psychoactive component of the plant Cannabis sativa, and endogenous mediators, such as anandamide, that belong to eicosanoid family.The biological effects of cannabinoids are mediated by a family of two G_i-coupled 7-transmembrane receptors, CB₁ and CB₂. The CB₁ receptor is found primarily in brain and mediates the psychoactive effects of cannabinoid ligands. The CB₂ receptor is expressed mainly in immune cells, including mast cells and CD40-activated B cells, where it mediates proliferation and inhibition of migration (Howlett et al., 2002). Activation of CB₂ inhibits the development of liver fibrosis (Julien et al., 2005). In bone, CB₂ is expressed in both osteoblasts and osteoclasts, and functions to prevent bone loss (Ofek et al., 2006). In addition, activation of CB₂ has an antinociceptive effect in animal models of neuropathic, inflammatory, and acute pain; this effect is mediated by release of endogenous opioids in the periphery (Ibrahim et al., 2005). Chemicon's cloned human CB₂-expressing cell line is made in the Chem-4 host, which supports high levels of recombinant CB₂ expression on the cell surface and contains high levels of promiscuous G proteins to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between CB₂ 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-4, an adherent cell line with no endogenous CB₂

2x10⁶ cells/vial

Chem-4

• CNR2
• CB-2
• OTTHUMP00000044841
• CB2
• CX5

Cells are frozen at 2 x 10⁶ cells/mL in DMEM/20% fetal bovine serum/100U/mL each penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

SPECIFICATIONS:
EC50 for calcium mobilization by CP55940: 78 nM
EC50 for calcium mobilization by WIN55, 212-2: 58 nM
EC50 for calcium mobilization by JWH 015: 603 nM.
Signal/noise ratio: 963

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)/ 0.5 mg/ml Hygromycin /100 U/ml each penicillin and streptomycin

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 growth media, and replace media after 8-24 h. Cells are typically passaged 1:10 with 0.05% Trypsin/0.2 g/L EDTA every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.