Details
The supplied Suc-LLVY-AMC is sufficient for 250 X 100 µl reactions.
Description
Proteasome Activity Fluorometric Assay Kit I is designed for assaying the proteasome’s chymotrypsin-like activity in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves Suc-LLVY-AMC and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively. The amount of the cell lysates or tissue extracts to be used in the reaction should be optimized.
AMC is used to produce a standard curve using a concentration range of 10-100 pmol.
Components
• AMC (0.1mM in DMSO)................................................30 µl
• 1000X SUC-LLVY-AMC (50 mM in DMSO)...................25 µl
• HEK293T cell lysates (5mg/ml).....................................100 µl
• MG-132..........................................................................1 mg
• MgCl2 (1M)....................................................................250 µl
• ATP (500 mM)................................................................100 µl
• DMSO............................................................................200 µl
• 20X Proteasome Assay Buffer.......................................1.35 ml