Figure 1.

Detailed procedure of bosutinib pull-down experiments. (A) Summary of bosutinib resin setups that were used for the investigation of affinity resin influences. In these experiments, the resin was incubated with 100 µL of K562 cell lysate (protein concentration of 1 µg/µL) at 4 °C for 2 h under continuous stirring (10 rpm). The elution volume was 250 µL, and the amount of trypsin used for protein digestion was 1.25 µg. For each affinity resin volume, the increasing concentration of bosutinib is marked by a shaded triangle. (B) Summary of the experimental conditions employed for the assessment of pull-down- and sample-processing effects. 50 nmol of bosutinib immobilized on 50 µL of the Sepharose resin (bosutinib concentration of 1 nmol/µL) was used for all listed pull-downs.