UbcH6, also known as UBE2E1 (GenBank Accession No. NM_003341), a.a. 2-end, with N-terminal His-tag, MW = 22 kDa, expressed in an E. coli cell expression system.
Immunogen Region
2-end
Tag
N-terminal His-tag
Formulation
40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 3 mM DTT, 20% glycerol, and 200 mM imidazole
Species
Human
Application
Useful in conjunction with E1 and E3 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Notes
50 ul reaction mix (4 mM ATP, 5 mM MgCl2, 1 mM DTT, UBE1 as an E1 (0.5 ug), UbcH6, Biotin-Ubiquitin (0.5 ug) and Smurf2 as an E3(2 ug) 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 (lambdaex=330 nm, lambdaem=620 nm).
Avoid freeze/thaw cycles.
GenBank
NM_003341
Mw(kda)
22 kDa
Synonyms
UbcH6, UBE2E1
Uniprot
P51965
Shipping Temperature
-80°C (dry ice)
Format
Aqueous buffer solution
Storage
At least 6 months at -80°C.
Reference
1. Sommer, T., et al. 1993. Nature 365 (6442):176-9. 2. Yang, M. et al. 1997. J. Biol. Chem. 272 :1970-1975.