ChemiScreen™ ChemR23 Calcium-Optimized FLIPR Cell Lines

ChemiScreen™ ChemR23 Calcium-Optimized FLIPR Cell Lines

• ChemiScreen
• Chemicon (Millipore)

2 vials

Full-length human ChemR23 cDNA

ChemR23 was discovered as an orphan receptor related to the chemoattractant receptors C3a, C5a and FPR1, and expressed on dendritic cells and macrophages (Samson et al., 1998). A ligand for ChemR23 was characterized as chemerin, a 15 kD proteolytically processed protein found in inflammatory sites; a 9 amino acid peptide from the C-terminus of chemerin is sufficient to activate ChemR23 (Wittamer et al., 2003, 2004). Chemerin expressed in lymphoid and microvascular endothelium mediates migration of ChemR23-expressing dendritic cells to lymphoid organs and vasculature at sites of inflammation (Vermi et al., 2005). In addition, a bioactive lipid, resolvin E1, was found to functionally interact with ChemR23 to reduce inflammation (Arita et al., 2005). Chemicon's cloned human ChemR23-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant ChemR23 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 ChemR23 and its ligands.

Calcium flux assay, ligand binding assays

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FUNCTION: SwissProt: Q99788 # Orphan receptor. Could be a chemotactic peptide receptor. May have a function in bone metabolism. Acts as a coreceptor for several SIV strains (SIVMAC316, SIVMAC239, SIVMACL7E-FR and SIVSM62A), as well as a primary HIV-1 strain (92UG024-2).
SIZE: 373 amino acids; 42322 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Prominently expressed in developing osseous and cartilaginous tissue. Also found in adult parathyroid glands.
DEVELOPMENTAL STAGE: Expressed during bone development.
SIMILARITY: SwissProt: Q99788 ## Belongs to the G-protein coupled receptor 1 family.

Human

SPECIFICATIONS: EC50 for calcium mobilization by Chemerin: ~ 4.7 nM

Chem-1

GPCR Cell Lines

Chemicon

Chem-1

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.

ChemR23 / CMKLR1

Q99788

peptide

Chemoattractant

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 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.

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A

1 mL per vial

• CMKLR1
• ChemR23
• DEZ
• MGC126106
• MGC126105
• CHEMR23

GPCRs

ChemiScreen™ ChemR23 Calcium-Optimized FLIPR Cell Lines

NM_004072.1

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)/100 U/ml each penicillin and streptomycin

2 x 10⁶ cells/mL