Chunk #15 — Experimental — Stable Expression of the A118/N40-hMOPR or G118/D40-hMOPR in CHO Cells — Metabolic Labeling with 35S-Labeled l-Methionine/l-Cysteine
CHO cells stably expressing 3HA tagged A118/N40- or G118/D40-hMOPR were washed with and preincubated in Dulbecco's modified Eagle's medium without l-methionine and l-cysteine for 1 h at 37°C. Pulse was performed in the fresh Met/Cys-free medium containing 150 μCi/ml of [35S]l-methionine/[35S]l-cysteine (1175.0 Ci/mmol). After 30-min incubation at 37°C, pulse was terminated by washing cells once with the chase medium (OPTI-MEM supplemented with 10% fetal bovine serum and 5 mM l-methionine), and chase was performed for indicated time periods.
