Source: Recombinant corresponding to aa130-end of human SMURF 2 with N-terminal FLAG tag expressed in a Baculovirus infected Sf9 cell expression system (NM_022739). Specific Activity: 0.00049 pmol/min/ug. Assay conditions: 50ul reaction mix (4mM ATP, 5mM MgCl2, 1mM DTT, E1 (0.5ug), E2 (2.8ug), Biotin-Ubiquitin (0.5) and Smurf2 (0-2ug) in 50mM Tris pH 7.4) incubate for 30min at 37C, then add to streptavidin coated plates and incubate for 30min at 37C. Add antibody against Smurf2, incubate 1 hour. Finally, incubate 1 hour with the Europeum-labeled secondary antibody. Time resolved fluorescence measured at RT (lambda-exc=330 nm, lambda-em=620 nm). Applications: Suitable for use in conjunction with E1 and E2 for the study of enzyme kinetics, screening inhibitors and selectivity profiling. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Storage and Stability: Aliquot to avoid repeated freezing and thawing. Store at -70C. Aliquots are stable for at least 6 months at -70C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.