Project description:Leukemic splenocytes from these commercial transgenic mice that developed fatal leukemia with massive splenomegaly were isolated at the time of the necropsy and subjected to gene expression profiling and phosphoprotein profiling in side by side comparison with CD22DE12-Tg BPL or CD22DE12_BCR-ABL double transgenic cells.
Project description:Phospho-peptide enrichment Protein extraction, reduction, alkylation and digestion were conducted as described above. Then, 15 mg TiO2-coupled beads were incubated with 500 μl Binding Buffer (BB) at RT for 10 min. Each aliquoted TiO2 was added into three 1.5 ml EP tubes, 5 mg for each and centrifuge 2,000 g for 2 min. The peptides were re-solved with 6μl BB and combined with 5 mg incubated TiO2 at RT for 30 min and centrifuge 1,000 g for 2 min. Repeat the solved procedures for the dried phosphopeptides twice with the 5 mg TiO2 and then discard the supernatant (Tong et al., 2019a).