Mpro Tracer
Tocris Bioscience, part of Bio-Techne | Catalog # 8126
Key Product Details
| Description: | SARS-CoV-2 main protease fluorescent ligand; TR-FRET acceptor |
| Purity: | ≥95% (HPLC) |
| Molecular Weight: | 909.41 |
Biological Activity for Mpro Tracer
Mpro Tracer is a high affinity (Kd = 40 nM) non-covalent SARS-CoV-2 main protease fluorescent ligand. Mpro Tracer can be used in fluorescent polarisation (FP) assay for inhibitor screening/characterisation. It can be used as a FRET acceptor in time-resolved fluorescence resonance energy-transfer (TR-FRET) experiments. Excitation and emission maxima (λ) are 492 nm and 518 nm.
Optical Data for Mpro Tracer
| Emission Color | Green |
|---|---|
| λabs | 492 nm |
| λem | 518 nm |
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Fluorescence polarisation assay to determine the inhibitory activity of nirmatrelvir on purified SARS-CoV-2 Main protease protein (Mpro) Mpro Tracer and purified Mpro were used at a concentration of 1 μM. A dose response over a range of inhibitor concentrations were measured. Assays were performed in 100 μL wells and left for 1 hour at room temperature before FP measurements were taken. Curve was plotted and fit using graphpad to obtain an IC50 value of 0.74 μM under the assay conditions. Data provided by the Hilgenfeld lab, Lubeck University. |
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3CLpro-C10778AA-FITC tracer HTRF assay The Kd of Mpro tracer was measured by varying the tracer concentration against a range of recombinant His6-tagged Mpro concentrations using an HTRF (TR-FRET) assay. MAb Anti-6HIS-Tb GOLD was used as the FRET donor and Mpro Tracer as the FRET acceptor (abs/em = 492nm/518nm). HTRF ratio (520/495) was determined using as Envision plate reader, measurement were recorded after 90 minutes. The Kd of Mpro Tracer was determined to be 40 nM. |
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