Fluorogenic, non-radioactive assay designed to measure chymotrypsin-like protease activity of purified 20S proteasome. This 96-well assay is useful for screening inhibitors and modulators of the 20S proteasome. The kit includes purified, human erythrocyte 20S proteasome (20µg). The Suc-LLVY-AMC fluorogenic peptide substrate supplied with this kit is a proteasome substrate commonly used for measuring chymotrypsin-like peptidase activity. The detection of proteolytic activity is based on the release of free AMC fluorophore and offers the following advantages: non-radioactive, convenient real-time kinetics of substrate cleavage, and excellent sensitivity. A proteasome inhibitor, epoxomicin, is included as a test inhibitor for screening purposes.
Fluorescence microscopy, HTS
Proteasome 20S (human) (purified)Suc-Leu-Leu-Val-Tyr-AMCProteasome Assay BufferSodium Dodecyl SulfateEpoxomicinAMC (7-Amino-4-methylcoumarin) Calibration Standard1/2 Volume White Microplates