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Applications: Study enzyme kinetics and screen small molecular inhibitors for drug discovery and High Throughput (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.A variant called B.1.1.529 (also known as the Omicron Variant) was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants. The 3CL protease of the Omicron variant is mutated at P132H compared to the wild-type SARS-CoV-2 strain.
Description: The 3CL Protease (B.1.1.529, Omicron Variant) (SARS-CoV-2) Assay Kit is designed to measure the activity of P132H mutated, Omicron variant 3CL Protease for screening and profiling applications, in a homogeneous assay with no time-consuming washing steps.The 3CL Protease Substrate is an internally quenched 14-mer fluorogenic (FRET) 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 glutamine and serine 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 comes in a convenient 384-well format, with purified 3CL Protease P132H (BPS Bioscience, #101328), fluorogenic substrate, and 3CL Protease assay buffer for 96 or 384 enzyme reactions. 3CL inhibitor GC376 is also included as a control.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Morse, J.S., et al., 2020 Chem.Bio.Chem. 21: 730 - 738.
2. Chi-Pang, C., et al., 2011 PLoS ONE 6(11): e27228.
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