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Applications: Study enzyme kinetics and screen small molecule inhibitors for drug discovery and High Throughput Screening (HTS) applications.
Background: Coronaviruses (CoVs) cause respiratory and intestinal infections in humans and animals. The 3CL protease, also known as Main Protease (Mpro), plays a vital role in processing the polyproteins that are translated from the viral RNA. Protease inhibitors that can block viral replication are promising potential drug candidates for the treatment of patients suffering from COVID-19 infection.T21I and A173V have been identified as mutations of interest for drug resistance.
Description: The 3CL Protease (T21I, A173V) (SARS-CoV-2) Assay Kit is a 96-well homogeneous fluorogenic assay designed to measure the activity of 3CL Protease mutated at T21I and A173V for screening and profiling applications, with no time-consuming washing steps. 3CL inhibitor GC376 is also included as a control.The 3CL Protease Substrate is an internally quenched 14-mer fluorogenic peptide (DABCYL-KTSAVLQSGFRKME-EDANS). When the donor (EDANS) and acceptor (DABCYL) fluorophores are in close proximity the energy emitted from EDANS is quenched by DABCYL (intact substrate). Upon proteolysis by 3CL, the peptide substrate is cleaved between the glutamine and serine residues to generate the highly fluorescent peptide fragment (SGFRKME-EDANS). The fluorescence intensity increases proportionally to the activity of 3CL. More information on the substrate, including MW and structure, can be found on our website (BPS Bioscience #79952).
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Supplied As: The kit contains enough purified 3CL Protease (T21I, A173V), fluorogenic substrate, and 3CL Protease assay buffer for 100 enzyme reactions.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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