| Assay Method Information | |
| | Confirmation of compounds inhibiting the binding between the RUNX1 Runt domain and CBFb-SMMHC via a fluorescence resonance energy transfer (FRET) assay |
| Description: | This assay is to identify inhibitors of the protein-protein interaction between the RUNX1 Runt domain and CBFbeta-SMMHC. This is accomplished by using a time resolved fluorescence resonance energy transfer (TR-FRET) assay. If an inhibitor is present, binding will not occur and the TR-FRET signal will be abated. The fluorescent CBF alpha/beta complex is incubated in black 384-well plate at room temperature for 1h. Positive control (PC) solution contained untagged CBFbeta-SMMHC. After incubation, The plate is read on a BMG Pherastar using a Lanthascreen filter set: 422ex, 530em and 480em. TR-FRET is determined as the ratio of 530/480 signals. This dose response assay was developed and performed to confirm hits originally identified in "uHTS identification of compounds inhibiting the binding between the RUNX1 Runt domain and CBFbeta-SMMHC via a fluorescence resonance energy transfer (FRET) assay" (AID 1434) and to study the structure-activity relationship on analogs of the confirmed hits |
| Affinity data for this assay | |
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