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Encompassing Amino Acids: 2-end
Applications: Useful in conjunction with E1 and E2 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 50 µl of reaction mixture 4 mM ATP, 5 mMMgCl2, 1 mM DTT, E1 (0.5 µg), E2 (2.8 µg),Biotinylated Ubiquitin (1 µg) in 50 mMTris pH 7.4 was incubated for 30 min at37°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.
Description: 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.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0,110 mM NaCl, 2.2 mM KCl, 80 µg/ml FLAGpeptide, 20% Glycerol and 3 mM DTT.
Genbank: NM_130838
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-FLAG-tags
Uniprot: Q05086
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 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).
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