| Bioactivity |
Specific Activity: 0.0027 pmol/min/µg. Assay conditions: 50 µl reaction mix (4mM ATP, 5mM MgCl2, 1mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotin-Ubiquitin (0.5 µg) and NEDD4 (0-2 µg) in 50mM Tris pH 7.4) incubate for 30min at 37°C, then add to streptavidin coated plates and incubate for 30min at 37°C. Add antibody against NEDD4, incubate 1h. Finally, incubate 1h with the Europeum-labeled secondary antibody. Time resolved fluorescence measured at room temperature (Lambda exc=330 nm, Lambda em=620 nm).
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