Mdm2 (mouse double minute 2, human homolog, also known as Hdm2), GenBank Accession No. NM_002392, (2-498) with N-terminal GST-tag, MW = 82 kDa, expressed in a baculovirus expression system.

Q00987-11

Mdm2,mouse double minute 2, human homolog, Hdm2

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 Mdm2 (0-0.5µg) 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-Mdm2 and HRP-labeled secondary antibody were incubated with the reaction mixture for 1 hour each, followed by chemiluminescent detection.

40 mM Tris pH 8.0, 110 mM NaCl, 2.2 mM KCl, 0.04% Tween-20, 20% glycerol and 3 mM DTT.

Aqueous buffer solution

>6 months at -80°C.

Useful in conjunction with E1 and E2 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

1. Fang, S., et al. (2000). J. Biol. Chem. 275, 8945-8951.
2. Chen, J et al. (1995). Mol. Med. 1, 142-152.

Avoid freeze/thaw cycles

Ubiquitin E3 ligase

Recombinant Protein