3CL Protease (T21I, T304I) (SARS-CoV-2) Assay Kit
The 3CL Protease (T21I, T304I) (SARS-CoV-2) Assay Kit is a 96-well homogeneous fluorogenic assay designed to measure the activity of T21I and T304I mutated 3CL Protease for screening and profiling applications, with no time-consuming washing steps. The kit contains enough purified 3CL Protease (T21I, T304I) (BPS Bioscience #101685), fluorogenic substrate, and 3CL Protease assay buffer for 100 enzyme reactions. 3CL inhibitor GC376 is also included as a control.
Figure 1: Illustration of the principle behind the 3CL protease assay.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).
Need us to run inhibitor screens or profile your compounds against 3CL Protease (T21I, T304I)? Check out our Protease Screening Services or Coronavirus Services.
Fluorescent microplate reader capable of reading λexc/λem=360 nm/460 nm
| Catalog # | Name | Amount | Storage |
| 101685 | 3CL Protease (T21I, T304I) (SARS-CoV-2)* | 2 µg | -80°C |
| 79952 | 3CL Protease Substrate (10mM) | 50 µl | -80°C |
| 79956 | 3CL Protease Assay Buffer | 25 ml | -20°C |
| 78013 | GC376, MW = 507.5** | 50 µg | -20°C |
| 0.5 M DTT | 200 µl | -20°C | |
| 79685 | Black, low binding microtiter plate | 1 | Room Temp |
* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.
**3CL inhibitor GC376 is provided as a control for 3CL inhibition.
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 T304I have been identified as mutations of interest for drug resistance.
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