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A live cell assay for SARS-CoV-2 3CL protease
Order 3CLglow Assay Kit

The 3CLglow Assay is a SARS-CoV-2 3CLpro inhibitor assay, a new technology developed by Montana Molecular in response to the COVID-19 pandemic. This assay provides a robust, biologically meaningful tool to evaluate therapeutic drugs that target SARS-CoV-2 replication.

  • 3C-like protease (3CLpro, Mpro, or Main protease) is essential for SARS-CoV-2 replication.
  • Discover new 3CL protease inhibitors.
  • A single BacMam delivers a gene encoding the green fluorescent 3CLglow Sensor as well as constitutively expressed red protein to check cell health.
  • BSL-1 kit is safe and easy to use.

Simple Assay Steps

  • Day 1 – Plate 35,000 HEK293 cells in each well.
  • Day 2 – Make transduction mix by combining 3CLglow BacMam, 3CLpro Enzyme, complete media, and sodium butyrate (included in 3CLglow Assay Kit).
  • Day 3 – Add inhibitor compounds and measure green and red fluorescence in either an endpoint assay to maximize throughput, or a kinetic assay for real-time detection of inhibitor action in living cells.

Images captured with a BioTek LionHeart FX automated microscope

3CLglow – Measure 3CLpro Inhibition

  • GC376 inhibits 3CLpro enzyme and is a reliable control for 3CLglow assay.
  • EC50 value for GC376 is consistent with reports by Vuong 2020 using live SARS-CoV-2 in culture.
  • Compatible with detection on fluorescence plate readers or imaging systems.

GC376 inhibits 3CLpro and is a reliable control for 3CLglow assays.

HEK 293 cells expressing both SARS-CoV-2 3CLprotease and fluorescent biosensor. Images captured on the BioTek LionHeart imaging plate reader.