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Encompassing Amino Acids: 150-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 mix (4 mM ATP, 5 mM MgCl2, 1 mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotin-Ubiquitin (0.5 µg) and SMURF1 (0-1.25µg) in 50 mM Tris pH 7.4) was incubated for 30 min at 37°C. The reaction mixture was transferred to a streptavidin-coated plate and incubated for additional 30 min at 37°C. Anti-SMURF1 and HRP-labeled secondary antibody were incubated with the reaction mixture for 1 hour each, followed by luminescent detection.
Description: Human SMURF1 (GenBank Accession No. NM_020429), a.a. 150-end, with N-terminal FLAG-tag, MW= 70 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 FLAG peptide, 3 mM DTT, 20% glycerol
Genbank: NM_020429
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal FLAG-tag
Uniprot: Q9HCE7
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Narimatsu M. et al., Cell 137 (2), 295-307 (2009)
2. Fukunaga E. et al., J. Biol. Chem. 283 (51), 35660-35667 (2008).
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