TUBEs: A Revolution in Polyubiquitin Isolation and Characterization -- The use of Tandem Ubiquitin Binding Entities (TUBEs) is emerging as an indispensable strategy for ubiquitin research. The first generation of these TUBEs bind K48- and K63-linked tetraUb chains with single digit nanomolar Kds, ~100 to 1000-fold more tightly than monomeric UBAs. TUBEs also protect proteins from DUBs and the proteasome, even in the absence of inhibitors normally required to block such activity. This allows efficient isolation of native polyUb chains and attached proteins from cell lines, tissues, and organs under conditions that are less likely to alter cell physiology than those listed above. TUBE1 and TUBE2 have been demonstrated to enrich for all polyUb chain linkage types, without discrimination, making these reagents appropriate even if the linkage type is not known for the protein of interest. K48 TUBE HF was developed to show enhanced selectivity for K48-linked polyubiquitin c
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Application: Protein Isolation, Protein Detection
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