| Solution Information | help | |
| Enzyme: | RNA-editing ligase 1, mitochondrial | |
| inhibitor: | BDBM38612 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine dose responses curves for compounds identified as active in a previous set of experiments entitled, "Fluorescence-based biochemical high throughput primary assay to identify inhibitors of Trypanosoma brucei RNA editing ligase 1 (TbREL1)" (PubChem AID 1117264) and "Fluorescence-based biochemical high throughput confirmation assay to identify inhibitors of Trypanosoma brucei RNA editing ligase 1 (TbREL1)" (PubChem AID 1117282). In this assay, REL1 enzyme is incubated with test compounds and fluorescently labeled RNA substrates. The assay substrate consists of a pair of RNA oligonucleotides, one with the FRET donor (6-FAM) and the other with FRET acceptor (CY5), which are annealed to a complementary strand. Uninhibited REL-1, in the presence of ATP, will ligate the RNA oligos with the FRET pair. STOP buffer is then added which denatures the annealed substrate and disrupts any unligated FRET pairings. As designed, test compounds t | |
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