"My laboratory has published two recent manuscripts reporting the use of FLAG-K63 and FLAG K48 TUBE reagents. We used these reagents to study the ubiquitination of the nonstructural 2 transmembrane protein of hepatitis C virus (HCV) by the cellular E3 ligase MARCH8. By being quite selective and easy to handle, these reagents helped us characterize the specific ubiquitin modification."
Dr. Shirit Einav, Associate Professor, Department of Microbiology and Immunology, Stanford University of Medicine
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
K48 TUBE HF was developed to show enhanced selectivity for K48-linked polyubiquitin chains (~20 nM) over all other linkages (>2 µM). It can be used alone or in conjunction with our other TUBE products, especially K63 TUBE and M1 (linear) TUBE to investigate polyubiquitin chain linkage in your substrate protein.
Note: The new K48 TUBE HF (UM607) has a comparable affinity as our existing K63 TUBE (UM604). Both UM607 and UM604 bind to their cognate Ub4s with a Kd of ~ 10-20 nM. Please note that the old K48 TUBE (UM605) binds K48-Ub4 with a Kd of ~200 nM.
Buffer: PBS, pH 7.2
Storage: Stable for up to 6 months at -80C. Aliquot reagent and avoid repeated freeze/thaw cycles.
- Far-Western detection of K48-polyubiquitinated proteins from cell and tissue extracts
- Isolation and enrichment of K48-polyubiquitinated proteins from cell and tissue extracts
- Isolate K48-polyubiquitinated proteins for proteomic studies
- 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
- Avoids overexpression of epitope-tagged ubiquitin for pull downs
- Compatible with FLAG® technology, providing flexibility and specificity
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|UM607_K48 TUBE HF FLAG_MSDS.pdf File Size 229.33KB Downloaded: 0|
|UM607_K48TUBE HF FLAG_Manual 2018.pdf File Size 225.47KB Downloaded: 0|