To isolate intracellular metabolites, cells were washed thoroughly
with PBS containing 0.05% glucose. Cells were lysed and Metabolism was
quenched by adding 5-10 ml of -80ÂșC methanol directly to the culture
dishes with attached cells. Lysates were collected, cell debris
removed by centrifugation and supernatants were frozen in liquid
nitrogen and lyophilyzed. Freeze-dried samples were processed
according to analytic requirements (see Maier et al., 2013)
and analysed by GC-MS, LC-MS or NMR. |