3CL Protease (B.1.1.529, Omicron Variant) (SARS-CoV-2) Assay Kit
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 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).
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 (B.1.1.529, Omicron Variant) (SARS-CoV-2)? Check out our Protease Screening Services or Coronavirus Services.
Fluorescent microplate reader capable of reading exc/em=360 nm/460 nm
96 Reactions
| Catalog # | Name | Amount | Storage |
| 101328 | 3CL Protease (B.1.1.529, Omicron Variant) (SARS-CoV-2)* | 5 µg | -80°C |
| 79952 | 3CL Protease Substrate (10 mM) | 25 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 |
| Plate sealing film | 1 |
384 reactions
| Catalog # | Name | Amount | Storage |
| 101328 | 3CL Protease (B.1.1.529, Omicron Variant) (SARS-CoV-2)* | 15 µg | -80°C |
| 79952 | 3CL Protease Substrate (10 mM) | 50 µl | -80°C |
| 79956 | 3CL Protease Assay Buffer | 25 ml | -20°C |
| 78013 | GC376, MW = 507.5** | 2 x 50 µg | -20°C |
| 0.5 M DTT | 200 µl | -20°C | |
| 79961 | Black, 384-well plate | 1 | Room Temp |
| Plate sealing film | 1 |
* 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. More information on GC376, including MW and molecular structure, can be found on our website (BPS Bioscience, #78013).
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.
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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