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.
The next generation of TUBEs: Linkage Specific Isolation of PolyUb Chains
Fluorescein-labeled (FLR) K48 TUBE HF binds K48-linked poly-ubiquitin with a 100-fold preference for K48 over other linkage types and provides a sensitive and cost-effective tool for determining the abundance of K48-linked polyubiquitin in the cell, or identifying the linkage-type associated with a particular protein of interest. FLR- K48 TUBE HF is useful for detection and characterization of K48-linked poly-ubiquitylated proteins by 'far Western' blotting and in situ detection (histo- or cyto-chemical staining).
FLR-K48 TUBE HF has one fluorescein molecule conjugated to a single, defined location within the protein.
Size: 50 µg
Buffer: PBS, pH 7.2
Storage: Stable for up to 6 months at -80C. Aliquot reagent and avoid repeated freeze/thaw cycles.
- "Far-Western" blot analysis
- Histo- or cyto-chemical staining. Sample fixation will require optimization.
- Nanomolar affinity for K48 poly-ubiquitin chains
- 100-fold preference for K48 poly-ubiquitin chains over all other linkages
- TUBEs display higher affinity towards polyubiquitins than most ubiquitin antibodies
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|UM507F_FLR-K48 TUBE HF_MSDS.pdf File Size 229.33KB Downloaded: 0|
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