ChemiScreen™ Human Recombinant CB₂ Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant CB₂ Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

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

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The cannabinoid delta-9-tetrahydrocannabinol is the principal psychoactive ingredient of marijuana. The proteins encoded by this gene and the cannabinoid receptor 1 (brain) (CNR1) gene have the characteristics of a guanine nucleotide-binding protein (G-protein)-coupled receptor for cannabinoids. They inhibit adenylate cyclase activity in a dose-dependent, stereoselective, and pertussis toxin-sensitive manner. These proteins have been found to be involved in the cannabinoid-induced CNS effects (including alterations in mood and cognition) experienced by users of marijuana. The cannabinoid receptors are members of family 1 of the G-protein-coupled receptors.

FUNCTION: SwissProt: P34972 # Involved in cannabinoid-induced CNS effects through G- protein mediated inhibition of adenylate cyclase. Could be a receptor for anandamide.
SIZE: 360 amino acids; 39681 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Not found in brain. Found in spleen.
SIMILARITY: SwissProt: P34972 ## Belongs to the G-protein coupled receptor 1 family.

Human

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

Chem-4

GPCR Cell Lines

Chemicon

Chem-4

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.

CB2

P34972

non-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 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.

Cannabinoid

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A

2x10⁶ cells/vial

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

GPCRs

ChemiScreen™ Human Recombinant CB₂ Calcium-Optimized Stable Cell Line

NM_001841.1