Human UBE3A (var1) (GenBank Accession No. NM_130838), a.a. 2-end, with N-terminal His-FLAG-tag, MW=99.8 kDa, expressed in a Baculovirus infected Sf9 cell expression system.

Q05086

UBE3A, E6AP

50 µl of reaction mixture 4 mM ATP, 5 mM MgCl₂, 1 mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotinylated Ubiquitin (1 µg) in 50 mM Tris pH 7.4 was incubated for 30 min at 37°C. Transfer the reaction mixture to a streptavidin-coated plate and incubate for 30 min at 37°C. Add antibody against UBE3A, incubate 1 hr. Then, add secondary HRP-labeled antibody and incubate 30 min. Finally, add HRP chemiluminescent substrates and read luminescence.

40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 80 µg/ml FLAG peptide, 20% 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. Dhe-Paganon S., et al. Biomol NMR. 51(1-2):185-90. (2011).
2. Jung EM, etal. Int J Mol Med. 28(5):697-704. (2011).

Avoid freeze/thaw cycles

Ubiquitin Activating Enzyme (E1)

Recombinant Protein