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[Cat.# F1100, F1101] MG-132

 

 

100 nM bovine 26S proteasome (Cat. # A1200) was incubated with DMSO or with 10 µM MG-132 for 10 min at 37 °C in 20 mM Tris, pH 7.1, 50 mM NaCl, 2 mM ATP, 5 mM MgCl2, 2 mM bME and 10% glycerol. The proteasome was then diluted 10X into 50 µM SUC-LLVY-AMC (Cat. # G1100) in a buffer containing 20 mM Tris, pH 7.1, 2 mM bME. The released AMC fluorescence was monitored by a plate reader.
 

Equal amount of whole cell lysates prepared from DMSO (lane 1) or 10 μM MG-132 (lane 2)-treated HEK293T cells were separated by SDS-PAGE and immunoblotted with an anti-Ub antibody. Cells were treated with DMSO or MG-132 for 4 hours.