Pe annexin 5 and 7 aad dual staining apoptosis detection kit

Apoptotic cells were detected using the PE-Annexin V and 7-AAD dual staining apoptosis detection kit (BD Pharmingen) by flow cytometry according to the manufacturer’s protocol. A total of 5 × 10⁵ MDA-MB-231 cells were seeded into a 100 mm sterile tissue culture Petri dish using 6 mL of DMEM/F-12. Then, cells were incubated at 37 °C in a 5% carbon dioxide atmosphere for 48 h. Subsequently, the medium was changed and cells were treated with different concentrations of drug solutions of 1 and 3 for 24 h. Cells were then harvested with cold 1× PBS containing 0.1 mM EDTA and subsequently washed twice with cold 1× PBS and finally resuspended in Annexin V binding buffer. Cells were then incubated with both Annexin V-PE and 7-AAD for 15 min in the dark at 25 °C. Data were analyzed in a BD Biosciences FACS Calibur flow cytometer within 1 h of sample preparation.

Apoptosis was detected using the PE-Annexin V and 7-AAD dual staining
apoptosis detection kit (BD Pharmingen) by flow cytometry according to the
manufacturer’s protocol. A total of 5 × 10⁵ MCF-7 cells
were seeded into a 35 mm sterile tissue culture Petri dish using 2 mL of DMEM.
Then, cells were incubated at 37 °C in a 5% carbon dioxide atmosphere for
another 48 h. Subsequently, the medium was changed and cells were treated with
different concentrations (IC₂₅ and IC₅₀) of oxamusplatin
for 24 h. Cells were then harvested with cold 1× PBS containing 0.1 mM
EDTA and subsequently washed with cold 1× PBS twice. The cells were
finally re-suspended in Annexin V binding buffer. Cells were then incubated with
both Annexin V-PE and 7-AAD for 15 min in the dark at 25 °C. Data were
analysed in a BD Biosciences FACS Verse flow cytometer within 1 h sample
preparation