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Article Novel iodoacetamido benzoheterocyclic derivatives with poten...

6.2.1. Cell lines and culture

The K562 human myeloid cell line was used in this study. K562 express the anti-apoptotic oncogene BCR–ABL and high levels of phosphorylated STAT5.

6.2.5. Flow cytometric evaluation of intracellular proteins

About 1 × 106 cells were washed twice with PBS (Sigma) and resuspended in 100 μL cytofix/cytoperm solution (Becton–Dickinson) at 4 °C. After 20 min the cells were washed twice with BD Perm/Wash™ buffer solution (Becton–Dickinson) and incubated with 20 μL of the specific fluorochrome(PE)-conjugated monoclonal antibody anti-p-STAT5 (Becton–Dickinson) at 4 °C. After 30 min the cells were washed twice and analyzed by flow cytometry. Alternatively, the cells were incubated with 2 μL PE-conjugated rat anti-mouse IgG1 monoclonal antibody (Becton–Dickinson) at 4 °C. After 30 min the cells were washed twice and analyzed by a FACScan flow cytometer (Becton–Dickinson).

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