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After this time, MeOH (81 L) was added and the RM was heated to 90 C for 1 h (for 15 ) and 14 h (for 16 and 17 )

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After mixing the sample or standard with ThioStar and incubating at room temperature for 15 minutes, the fluorescent product is read at 510 nm in a fluorescent plate reader with excitation at 390 nm

As with older cellular proteomics studies, some secretomics approaches employ SDS-PAGE for separation and in-gel digestion to acquire peptides for MS (Athenstaedt et al., 2006