"Total of 10 mg of thiol alkylated protein mixture (15 N/14 N) was loaded onto one 190 mm×180 mm preparative SDS-PAGE slab gel (10%). A small fraction of slab gel was sliced vertically and stained with Coomassie brilliant blue to examine the quality of electrophoresis separation of sample proteins. The rest part was sliced horizontally and evenly into 10 strips and further diced into 1-mm 3 cubes. The in- gel trypsin digestion was performed as previously described. The digested peptides were extracted from the gel cubes by 1% FA in 50% ACN for 3 times. The ACN solvent in the combined peptide extract in each falcon tube was vaporized, lyophilized to a complete dry powder and then stored at -80°C.
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Citation: Hu Q, Guo G, Yang Z, Li Y, Xia Y (2014) Stable Isotope Metabolic Labeling-Based Quantitative Thiol Redox Proteomic Analysis of Hydrogen Peroxide-treated Arabidopsis plant. J Proteomics Bioinform 7: 121-133."
Last date updated on November, 2024