| Assay Method Information | |
| | Fluorescence Cell-Free Homogeneous Counterscreen to Identify Inhibitors of the RanGTP-Importin-beta complex. |
| Description: | Keywords: Counterscreen, FRET assay, HTS, YIC probe, Dose response, Assay Overview: This is a fluorescence resonance energy transfer (FRET)-based biochemical assay used as counterscreen for this project. YIC probe is a protein containing the Importin-beta binding domain of Importin-alpha flanked by cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP). This probe undergoes robust intramolecular FRET that should not be affected by specific hits from primary screen. In this assay, we are looking for compounds do not inhibit FRET signal from YIC probe. The inhibiting potency is assigned by its IC50, Expected Outcome: Active compound will show a reduction of fluorescent ratio (IYFP/ICFP) in a concentration-dependent manner. |
| Affinity data for this assay | |
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