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| | Fluorescence Binding Assay |
| Description: | The fluorescence buffer was degassed and aerated with pure nitrogen gas to remove dissolved oxygen. The assay was carried out on a FluoroMax-2 fluorometer. The excitation and emission wavelengths were 284 nm and 341 nm, respectively. The inhibitor solution was titrated into EGFR kinase solution, and the emission fluorescence intensity was read after addition of inhibitor, and the average of five measurements was recorded. A blank assay was performed in exactly the same manner except that the buffer without inhibitor was used for the titration. Dissociation constants (Kd) were determined by nonlinear fitting of the fluorescence data using a modified static quenching model. |
| Affinity data for this assay | |
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