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Mistake #6: Temperature Cycling

In brief, after labeling and washing, cells were lysed with radioimmunoprecipitation assay (RIPA) lysis buffer supplemented with 1 protease inhibitor cocktail (Thermo Scientific, 78429)

Quantitative profiling using HPLC-MS/MS is highly reliable when employing an isotope dilution approach, wherein stable isotope-labeled internal standards (IS) effectively compensate for matrix effects (examples are shown for GSH and Glu in Fig