| Solution Information | help | |
| Enzyme: | Protein Rev | |
| inhibitor: | BDBM52490 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this counterscreen assay was to determine whether compounds identified as active in a set of previous experiments entitled, "Fluorescence polarization-based primary biochemical high throughput screening assay to identify inhibitors of gld-1 protein - TGE RNA interaction" (AID 2280), were non-selective or assay artifacts due to inhibition of the interaction between HIV Rev peptide and RRE RNA. This biochemical assay is based on the ability of the Rev peptide to form a complex with the Rev-Responsive Element (RRE) of double-stranded viral mRNA. Both binding partners have been labeled with appropriate fluorophores in order to monitor their interaction by fluorescence resonance energy transfer (FRET). A Cy5-labeled RNA target harboring the Rev Response Element (Cy5-RRE) is first incubated with test compounds, followed by addition of a Fluorescein-labeled Rev peptide (Fl-Rev) and reading of the FRET fluorescence. A high FRET is indicative of proper binding o | |
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