ChemiScreen™ Human Recombinant GPR41 Free Fatty Acid Receptor Calcium-Optimized Stable Cell Line

ChemiScreen™ Human Recombinant GPR41 Free Fatty Acid Receptor Calcium-Optimized Stable Cell Line

• ChemiScreen
• Chemicon (Millipore)

2 vials

GPR41 is a GPCR that, along with GPR43, is activated by short chain carboxylic acids formate, acetate, proprionate, butyrate and pentanoate (Brown et al., 2003; Brown et al., 2005). Binding of these ligands to GPR41 selectively activates Gi to inhibit cAMP accumulation. Expression of GPR41 is prominent in adipose tissue, increases during differentiation of cultured adipocytes, and allows short chain carboxylic acids to stimulate leptin synthesis in adipocytes (Xiong et al., 2003). Chemicon's cloned human GPR41-expressing cell line is made in the Chem-4 host, which supports high levels of recombinant GPR41 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 GPR41 and its ligands.

Calcium flux assay, ligand binding assays

Human

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FUNCTION: SwissProt: O14843 # Receptor for short chain fatty acids through a G(i)- protein-mediated inhibition of adenylyl cyclase and elevation of intracellular calcium. The rank order of potency for agonists of this receptor is propionate = pentanoate = butyrate > acetate > formate.
SIZE: 346 amino acids; 38649 Da
SUBCELLULAR LOCATION: Cell membrane; Multi-pass membrane protein.
TISSUE SPECIFICITY: Highest level in adipose tissue, and lower expression across all tissues tested.
SIMILARITY: SwissProt: O14843 ## Belongs to the G-protein coupled receptor 1 family.

Human

EC50 for calcium mobilization by Sodium Propionate: ~ 16 uM

TRANSFECTION: Full-length human GPR41 cDNA

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 Hygromycin (500 ug/ml) Geneticin (G418)/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/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.

FFA3 / GPR41

O14843

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 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.

Free Fatty Acid

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A

2 x 10⁶ cells/vial

• FFAR3
• GPR41
• FFA3R

GPCRs

ChemiScreen™ Human Recombinant GPR41 Free Fatty Acid Receptor Calcium-Optimized Stable Cell Line

NM_005304.2