| Assay Method Information | |
| | Ebola Antiviral Assay (EBOV) |
| Description: | HEp-2 cells were plated in 96-well plates at the density of 40,000 cells/well. On the next day, modified vaccinia virus Ankara-T7 (MVA-T7) at the multiplicity of infection of 1 was added to provide T7 RNA polymerase. After 2 hours of viral transduction, each well was transfected with Lipofectamine2000 (Thermo Fisher) with 0.01 g mixture of 6 plasmids including Ebola minigenome, plasmids encoding Ebola L, NP, VP-35, VP-30 proteins. After 48 hours of further incubation, cells were lysed with RIPA buffer (Pierce), transferred to a black 96-well plate and the fluorescence was read at 0.1 sec/well at ex485 nm, emission 535 nm on a Victor plate reader. Sigmoidal dose-response curves used to generate 50% inhibitory or effective concentrations were analyzed by nonlinear regression using the four-parameter logistic equation (GraphPad Prism). |
| Affinity data for this assay | |
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