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| | In vitro EGFR TK Inhibition Assays |
| Description: | The experiments were performed according to the instructions of the manufacturer. The EGFR TK activity was performed using Kinase-Glo Plus luminescence kinase assay kit (Promega). It measures kinase activity by quantitating the amount of ATP remaining in solution following a kinase reaction. The luminescent signal from the assay is correlated with the amount of ATP present and is inversely correlated with the amount of kinase activity. Briefly, 96-well plates were precoated with EGFR, EGF, and tested compound in a ratio of 2:2:1 that were added.The reaction mixtures were incubated for 40 min at room temperature while being shaken. After completion of the reaction, 20 µL of ADP-GloTM Plus Luminescence kinase assay solution were added, incubate the plate for 30 min at room temperature. The corrected activity for each protein kinase target was determined by removing the blank control value. Luminescence signal was measured using aTecan Infinite Spectramax M2e microplate reader. |
| Affinity data for this assay | |
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