cbil.upenn.edu:RAD.Protocol:1545:Protocol - cbil.upenn.edu:RAD.Protocol:1545:Protocol
Cells were grown in P-100 tissue culture plates. Growth Medium RPMI-1640 was supplemented with 10% fetal calf serum, 10mM Hepes, 2mM L-glutamine, 1mM Sodium-pyruvate, and 0.05 mM 2-mercaptoethanol. Growth medium was changed every 2-3 days. This medium was changed to medium containing 1mM oleate/palmitate (O/P) at a given time point before the cells reached confluency. The O/P Medium was prepared from a 10mM oleate/ palmitate stock solution prepared in 1% BSA (fatty acid free). Oleate/palmitate were mixed at a 2:1 molar ratio.
Time Medium was Changed Prior to Confluency