3CL Protease, MBP-tagged (SARS-CoV-2) Assay Kit

The3CL Protease Assay Kitis designed to measure 3CL Protease activity for screening and profiling applications, in a homogeneous assay with no time-consuming washing steps. The kit comes in a convenient 96-well format, with purified 3CL Protease MBP-tag (BPS Bioscience #100707), fluorogenic substrate, and 3CL Protease assay buffer for 100 enzyme reactions. 3CL inhibitor GC376 is also included as 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 glutamine and serine by the 3CL protease 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) .The 3CL Protease, Untagged (SARS-CoV-2) Assay Kit (BPS Bioscience #78042) demonstrates better sensitivity than the 3CL Protease, MBP-tagged (SARS-CoV-2) Assay Kit, therefore we highly recommend using #78042 for best results.Figure 1: Illustration of the principle behind the3CL 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 (SARS-CoV-2) ? Check out ourProtease Screening ServicesorCoronavirus Services.This product has beencited 57 times.

Product Specifications

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.

Applications

Great for studying enzyme kinetics and for high throughput (HTS) applications.

Format

96 Reactions:Catalog #ComponentAmountStorage100707Recombinant 3CL Protease, MBP-tag*20 µg-80°CAvoidfreeze/thawcycles!799523CL Protease Substrate (10 mM) 25 µl-80°C799563CL Protease Assay Buffer25 ml-20°C78013GC376, MW=507.550 µg-20°C0.5 M DTT200 µl-20°C79685Black, low binding microtiter plate1RoomTempPlate sealing film1*The exact concentration of protein is lot-specific and will be indicated on the tube containing the protein.384 Reactions:Catalog #ComponentAmountStorage100707Recombinant 3CL Protease, MBP-tag*60 µg-80°CAvoidfreeze/thawcycles!79952-13CL Protease Substrate (10 mM) 50 µl-80°C799563CL Protease Assay Buffer25 ml-20°C78013GC376, MW=507.550 µg-20°C0.5 M DTT200 µl-20°C79961384-well black, low binding microtiter plate1RoomTempPlate sealing film*The exact concentration of protein is lot-specific and will be indicated on the tube containing the protein.