Assay Method Information

Assay Name:  ABL1 (T315I) Kinase Activity Assay
Description:  Serially diluting the compound of the present invention from 1 uM initial concentration in three-fold fashion and formulating 10 concentrations (50.8 pM, 152.0 pM, 457.0 pM, 1.37 nM, 4.12 nM, 12.3 nM, 37.0 nM, 111.0 nM, 333.0 nM and 1.0 uM). 5.0 uM Abltide was added into each well and then human T315I mutant enzyme was added. [gamma-33P] ATP was added at room temperature, with final concentration of 1.0 uM, and the reaction was performed for 120 minutes. 20 ul aliquots were transferred onto the ion exchange chromatography paper P81. The paper was thoroughly washed with a 0.75% phosphoric acid solution three times, and then washed with acetone once. Finally, gamma-33P radioactivity was measured.
Affinity data for this assay
 

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