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Encompassing Amino Acids: 2-170(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 (0.5 µg), UEV1A or other E2, Biotin-Ubiquitin (0.5 µg) and Smurf2 (2 µg) in 50 mM Tris, pH 7.4) for 30 min at 37°C. Add to Streptavidin-coated plates for 30 min at 37°C. Incubate with anti-Smurf2, then HRP-labeled secondary antibody, and chemiluminescent detection.
Description: UEV1A (ubiquitin-conjugating enzyme E2 variant 1), also known as UBE2V1 or CROC-1 (GenBank Accession No. NM_021988), a.a. 2-170(end), with N-terminal FLAG-tag, MW=20 kDa, expressed in E. coli.
Format: Aqueous buffer solution
Formulation: 45 mM Tris-HCl, pH 8.0, 124 mM NaCl, 2.4 mM KCl, 90 µg/ml Flag peptide, 10% glycerol, and 3 mM DTT.
Genbank: NM_021988
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal FLAG-tag
Uniprot: Q13404
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Thomson TM, et al. (1998). FEBS Lett. 423
(1): 49-52.
2. Hau DD, et al. (2006). Biochemistry 45
(32): 9866-77.
3. Ma L, et al. (1998). Oncogene 17 (10):
1321-6.
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