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

TheUntagged 3CL 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 untagged 3CL Protease (BPS Bioscience #100823), fluorogenic substrate, and 3CL Protease assay buffer for 100 enzyme reactions. 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 glutamine and serine by the 3CL protease to generate the highly fluorescent peptide fragment (SGFRKME-EDANS) . EDANS has an excitation peak at 336 nm and an emission peak at 455 nm. 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 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) .Assay PrincipleNeed us to run inhibitor screens or profile your compounds against 3CL Protease, Untagged (SARS-CoV-2) ? Check out ourProtease Screening ServicesorCoronavirus Services.This product has beencited 45 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 #ComponentAmountStorage1008233CL Protease (SARS-CoV-2), no tag*>2 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.Excess material is provided for ease of retrieval.384 Reactions:Catalog #ComponentAmountStorage1008233CL Protease (SARS-CoV-2), no tag6 µg-80°CAvoidfreeze/thawcycles!799523CL Protease Substrate (10 mM) 50 µl-80°C799563CL Protease Assay Buffer25 ml-20°C78013GC376, MW=507.52 x 50 µg20°C0.5 M DTT200 µl-20°C79961Black, 384-well plate1RoomTempPlate sealing film1*The exact concentration of protein is lot-specific and will be indicated on the tube containing the protein.