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Coomassie-stained urea SDS-PAGE gel demonstrating the formation of homogeneously K11-linked Ub4 via a 20-hour incubation of Alloc-protected middle-15N-labeled K11-linked Ub3-SR and TFA-treated, Alloc-protected K11Boc Ub monomer (TFA K11Boc UbA). The contaminating Ub2 band from an impure sample of K11Boc is inert as it is visible both before and after the ligation reaction. The sample after ligation was loaded at higher concentration to emphasize the formation of Ub4.
