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Encompassing Amino Acids: 2-end
Applications: Useful in conjunction with E1 and E3 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: 50 µl reaction mix (4 mM ATP, 5 mM MgCl2, 1 mM DTT, UBE1 as an E1 (0.5 µg), UbcH7, Biotin-Ubiquitin (0.5 µg) and Smurf2 as an E3(2 µg) in 50 mM Tris pH 7.4) for 30 min at 37°C, then add to streptavidin coated plates for 30 min. at 37°C. Incubate with anti-Smurf2, thenEuropium-labeled secondary antibody, and detect by time resolved fluorescence (λex=330 nm, λem=620 nm).
Description: UbcH7, also known as UBE2L3 (GenBank Accession No. NM_003347), a.a. 2-end, with N-terminal His-tag, MW = 18 kDa, expressed in an E. coli cell expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 3 mM DTT, 20% glycerol, and 200 mM imidazole
Genbank: NM_003347
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal His-tag
Uniprot: P68036
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Katsanis, N., et al. 1998. Genomics 51: 128- 131.2. Cook, W.J. et al., 1997. Biochemistry 36 (7): 1621-1627.
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