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Assay Method Information

Assay Name:  Confirmation of compounds inhibiting the binding between the RUNX1 Runt domain and CBFb-SMMHC via a time resolved fluorescence resonance energy transfer (TR-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: 337ex, 520em and 490em. TR-FRET is determined as the ratio of 520/490 signals. This dose response assay is developed and performed to confirm hits originally identified in "Identification of compounds inhibiting the binding between the RUNX1 Runt domain and CBFbeta-SMMHC via a time resolved fluorescence resonance energy transfer (TR-FRET) assay"(AID 1438) and to study the structure-activity relationship on analogs of the conf
Affinity data for this assay
 

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Last update November 1, 2007
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