Assay Method Information

Assay Name:  scanning fluorimetry assay of Vero E6 cells
Description:  The differential scanning fluorimetry (DSF) assay was carried out on a CFX96 real-time PCR Detection System (Bio-Rad). All the experiments were performed in the following buffer: 20 mM HEPES pH 6.5 containing 120 mM NaCl. The Mpro protein (final concentration: 2 μM) was pre-incubated with 40 μM of compounds for 30 min. 5×SYPRO Orange dye (Sigma) was added to probe the thermal denaturation from 20°C to 95°C at a scan rate of 1.5 °C/min. The melt temperature (Tm) was calculated by using a Boltzmann model in GraphPad Prism 8.0 software. The thermal shift (ΔTm) was calculated using the equation ΔTm=Tm(compound)-Tm(DMSO). All experiments were performed in triplicate, and the values are presented as mean ± SD. Vero E6 cells are used.
Affinity data for this assay
 

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