Figure 3.

Retrieval of DUBs from whole cell lysate using the catch-and-release probe.

A) 293T cells were grown overnight in the presence of [³⁵S]cysteine/ methionine. Cells were lysed in NP40 lysis buffer, precleared with streptavidin beads, followed by incubation with 1 μM of the indicated probe at 4°C. Labeled product was retrieved with streptavidin agarose, eluted with the relevant elution buffer, and analyzed by SDS-PAGE.

B) 293T cells were transiently transfected with YOD1 and treated as in A), though exclusively with the LEV probe. Where indicated, the eluate was redissolved in NP40 buffer, followed by immuno-retrieval of YOD1.