A kinase profiler service from Millipore (Billerica, MA) was used to assess the direct inhibitory effect of Sb-EE against Src, Syk and IRAK1. An Sb-EE stock solution was prepared to 50× final assay concentration in 100% DMSO. The reaction was initiated by incubating with a Mg/ATP mixture. After 40 min at room temperature, the reaction was stopped by adding 0.5% phosphoric acid. Then, 10 μL of the reaction solution was spotted onto a P30 filtermat and washed four times for 4 min in 0.425% phosphoric acid, followed by one wash in methanol. After this, the sample was dried and by scintillation counts.

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