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Schematic presentation of the method. N‐ethylmaleimide (NEM), an alkylating agent, forms stable bonds with cysteine and other thiols in cytosol, enabling them to be permanently blocked and prevent disulfide bond formation, while leaving cystine in lysosomes intact. This process is then followed by cell lysis and acid precipitation of proteins. This causes cystine to be present in the acid‐soluble fraction, which can be quantified by HPLC‐MS/MS
