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Synthesis of ubiquitin-αMeCys and model ligation. a Analytical HPLC (RP-C18) of crude and purified Ub-αMeCys. MALDI-TOF MS, m/z = 8682 [M + H]+, calc. Av. = 8682. b Model ligation of NEMO298–308hdmb with Ub-MES thioester, time-course monitoring of ligation reaction by analytical HPLC, RP-diphenyl. Bottom, analytical HPLC of NEMO298–308-Ub (after hdmb removal) (d) and MALDI-TOF MS, m/z = 9818 [M + H]+, calc. Av. = 9817
