Human SMURF2 (GenBank Accession No. NM_022739), a.a. 130-end, with N-terminal FLAG-tag, MW= 72 kDa, expressed in a Baculovirus infected Sf9 cell expression system.

Q9HAU4

SMAD specific E3 ubiquitin protein ligase 2, SMURF2

50 µl reaction mix (4 mM ATP, 5 mM MgCl₂, 1 mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotin-Ubiquitin (0.5 µg) and SMURF2 (0-2 µg) in 50 mM Tris pH 7.4) incubate for 30 min. at 37°C, then add to streptavidin coated plates and incubate for 30 min at 37°C. Add antibody against SMURF2, incubate 1 hr. Finally, incubate 1 hr. with the Europium-labeled secondary antibody. Time resolved fluorescence measured at room temperature (λexc=330 nm, λem=620 nm).

45 mM Tris-HCl, pH 8.0, 124 mM NaCl, 2.4 mM KCl, 90 ?g/ml FLAG peptide, 10% glycerol, and 3 mM DTT.

Aqueous buffer solution

>6 months at -80°C.

Useful in conjunction with E1 and E2 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

1. Narimatsu M. et al., Cell 137 (2), 295-307 (2009)
2. Fukunaga E. et al., J. Biol. Chem. 283 (51), 35660-35667 (2008).

Avoid freeze/thaw cycles

Ubiquitin E3 ligase

Recombinant Protein