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Gel lanes were cut in several pieces, and proteins were reduced, alkylated, and digested by overnight incubation with trypsin (bovine sequencing grade, Roche Applied Science, Monza, IT) according to published protocols ( 2.10 Identification of selected antigen by LC-MALDI Protein immunopurifications were precipitated in ice-cold acetone overnight and resuspended in 40 µL of NH 4 HCO 3 50 mM, urea 2 M solution, and digested with 5 µL of trypsin solution (0.2 µg/µL) overnight at 37°C