Ubiquitin vinyl sulfone, (HA-tag)

As indicated on product label or CoA when stored as recommended. Stable for at least 6 months after receipt when stored at -80°C. Once reconstituted store HA-Ub-VS solution at -80°C, avoid multiple freeze/thaw cycles and use within 1 month.

Typical assay set-up: HA-Ub-VS probe concentration: 0.5-5.0μM. Purified enzyme concentration: 0.1-1.0μM; Buffer: 50mM TRIS, pH7.5, 150mM NaCl, 1mM DTT. Incubate at 37°C for 30 minutes. Modification of DUB enzymes by site-directed HA-Ub-VS probes detected by Western blotting with either HA antibody (general reactivity) or an antibody specific to particular DUB of interest. Note: When probing complex solutions such as cell lysates the optimal amount of total sample protein required will depend on the specific lysate to be analyzed and must therefore be determined for the individual lysate of interest.

Suggested uses: 1. Deubiquitinylating enzyme active site-directed probes for detection of novel DUBs. 2. Determination of deubiquitinylating enzyme activity of putative DUBs. 3. Identification of DUBs from biological samples via immunoprecipitation and proteomic methods. 4. Characterization of deconjugating enzymes for Ub/Ubl specificity.