SMURF2, N-terminal FLAG-Tag, Recombinant, Human (Smad Ubiquitination Regulatory Factor 2, E3 Ubiquitin-Protein Ligase SMURF2, Smad-Specific E3 Ubiquitin Ligase 2)

Dry Ice

72

Supplied as a liquid in 25mM Tris-HCl, pH 8.0, 100mM sodium chloride, 0.05% Tween 20, 100ug/ml FLAG peptide, 3mM DTT, 20% glycerol.

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 37ºC, then add to streptavidin coated plates and incubate for 30min at 37ºC. 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 -70°C. Aliquots are stable for at least 6 months at -70°C. 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.