Table 7.
List of flow cytometry analyses and staining.
| Analyses | Staining Solution | Company |
|---|---|---|
| Cell cycle * | 10% Triton X-100 0.5 mg/mL ribonuclease A 0.025 mg/mL propidium iodide–PI In 500 µL PBS |
Sigma-Aldrich, Steinheim, Germany |
| Apoptosis | Annexin V-Alexa Fluor 647 1:100 (catalogue no. A23204) | Thermo Scientific, Rockford, IL, USA |
| PI (5 mg/mL) 1:500 | Sigma-Aldrich | |
| Mitochondrial membrane potential | TMRE (tetramethylrhodamine ethyl ester per chlorate) final conc. 0.1 µM | Molecular Probes, Eugene, OR, USA |
| Caspase activation | Cleaved caspase-3 rabbit mAb PE conjugate 1:100 (#9978) | Cell Signaling, Danvers, MA, USA |
| Cleaved caspase-7 rabbit mAb PE conjugate 1:100 (#42542) | ||
| Proteins analysis | Cleaved PARP rabbit mAb PE conjugate 1:100 (#8978) | |
| Bcl-2 mouse mAb PE conjugated 1:100 (#26295) | ||
| Phospho-Bcl-2 (Ser 70) rabbit mAb Alexa Fluor 488 conjugate 1:200 (#2834) |
* After harvesting, cell suspension is fixed in cold 70% ethanol and kept at −20 °C overnight.