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Generation of branched warhead-containing triUb probe for pulldown and branched triUb for binding assessment. A. Synthesis of triUb-NC(1,1)-PA2 probes via ligation of HA-Ub1-75-NC and Ub1-75-PA with a double-cysteine mutation introduced at requi-site lysine residues. B. Synthesis of triUb-NC(1,0) via ligation of enzymatically generated diUb-D77 with Ub1-75-NC.
