Solution Information | help | |
Enzyme: | Metabotropic glutamate receptor 4 [Q124R] | |
inhibitor: | BDBM55090 | |
substrate: | n/a | |
Solution Type: | Aqueous | |
pH at Preparation: | n/a | |
Temp. Prep.: | n/a | |
Comments: | Cell culture, plating, and dye loading. HEK/GIRK cells stably expressing the M4 muscarinic receptor were grown in 45% Dulbecco's Modified Eagle Media (DMEM), 45% Ham's F12, 10% fetal bovine serum (FBS), 100 units/ml penicillin/streptomycin, 20 mM HEPES (pH 7.3), 1 mM sodium pyruvate, 2 mM glutamine, and 700 ug/ml G418. The rat mGluR4 cell line was prepared by PCR amplification of the entire coding sequence of each receptor and cloning into pIRES puro 3 (Invitrogen). Cloning sites were BamHI/Not I. HEK/GIRK/M4 cells were transfected with 24 ug of DNA and stable transfectants were selected with puromycin. And a monoclonal cell lines was established. Cells were grown in 45% Dulbecco's Modified Eagle Media (DMEM), 45% Ham's F12, 10% fetal bovine serum (FBS), 100 units/ml penicillin/streptomycin, 20 mM HEPES (pH 7.3), 1 mM sodium pyruvate, and 2 mM glutamine (Growth Media. mGluR/GIRK lines were supplemented with 600 ng/ml puromycin dihydrochloride (Sigma-Aldrich) and 700 ug | |
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