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[Cat.# F1300, F1301] Carfilzomib (PR-171)

 

 

100 nM bovine 26S proteasome (Cat. # A1200) was incubated with DMSO or with 10 mM Carfilzomib (PR-171) 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 mM 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. 26S proteasome + DMSO (open circle); 26S proteasome + Carfilzomib (PR-171) (solid circle).
 

Equal amount of whole cell lysates prepared from DMSO (lane 1) or 10 μM Carfilzomib (PR-171) (lane 2)-treated HeLa cells were separated by SDS-PAGE and immunoblotted with an anti-Ub antibody. Cells were treated with DMSO or Carfilzomib (PR-171)1 for 16 hours.