Annexin 5 fitc and propidium iodide double staining kit

Apoptotic cells were analyzed by flow cytometry with an Annexin V‑FITC and propidium iodide (PI) double-staining kit (BD Biosciences). Cells used for flow cytometry were collected by trypsinization and resuspended in 100 µl 1X binding buffer containing Annexin V and PI. Cells were stained for 15 min at 25°C in the dark before being analyzed by flow cytometry. All the cytometry data in the present study were acquired with a BD FACSVerse flow cytometer and analyzed using FlowJo software (version 10.0.7; Tree Star, Inc.).

Apoptotic K562 cells were measured using Annexin V‐FITC and propidium iodide (PI) double‐staining Kit (BD Biosciences, Franklin Lakes, NJ, USA) according to the manufacturer's instructions. Briefly, 5.0 × 10⁵ cells were washed with ice‐cold PBS, resuspended in 195 μL binding buffer, and stained for 10 minutes at room temperature with 5 μL FITC conjugated anti‐Annexin V antibody. Unbound Annexin V antibody was removed by washing with binding buffer. Percentage of apoptotic K562 cells (Annexin V positive) was determined by flow cytometry analysis. Flow cytometry was carried out using a FACSCalibur cytometer (BD Biosciences) and analyzed using CellQuest software (BD Biosciences).