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Left: SDS-PAGE analysis of the assembly and purification of polyubiquitin chains for FRET-based studies, which consist of a proximal Cy3-labeled ubiquitin and several Cy5-labeled distal ubiquitins. Double-labeled chains were visualized by Cy3 fluorescence (magenta) and Cy5 fluorescence (cyan). Right: the crystal structure of a K48-linked ubiquitin dimer (PDB ID: 1AAR) shows the distance between the Cy3-donor labeled N terminus of the first ubiquitin in the chain and the Cy5-acceptor-labeled N terminus of the second ubiquitin in the chain.

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