Annexin 5 fluorescein isothiocyanate fitc apoptosis kit

Cell apoptosis was detected using Annexin V-fluorescein isothiocyanate (FITC) Apoptosis Kit (Biovision, Milpitas, CA) at 48 hours post-transfection. Cells (2 × 10⁵) for each reaction were washed in phosphate-buffered saline (PBS) for 2 times, after which 100 μL Binding Buffer and 2 μL Annexin-V FITC (20 μg/mL) were added, and then the cells were incubated in the dark on ice for 15 minutes. After the incubation, 400 μL PBS and 1 μL propidium iodide (PI) were added and the cells were immediately analyzed by flow cytometry BD FACSCalibur (BD Biosciences, San Jose, CA). Cells in the lower right quadrant (FITC positive and PI negative) were considered to be apoptotic cells.

The different stages of apoptotic cells were examined using the Annexin V-Fluorescein Isothiocyanate (FITC) Apoptosis Kit (BioVision, Milpitas, CA, USA). Early and late stages of apoptotic cells were stained with Annexin V, and necrotic cells were stained with propidium iodide and then analyzed using the FACSVerse flow cytometer (BD Biosciences, San Jose, CA, USA).

The apoptosis status of the cells in each group was determined by an Annexin V-fluorescein isothiocyanate (FITC) Apoptosis kit (BioVision, Inc., Milpitas, CA, USA), according to the manufacturer's protocols. Briefly, floating and trypsinized adherent cells (5×10⁵) from each group were collected and re suspended in 500 µl PBS containing 5 µl Annexin V-FITC and 5 µl PI, and incubated for 5 min in the dark at room temperature. Analysis was subsequently performed using a flow cytometer (BD Biosciences). Cell Quest Pro software (BD Biosciences) was used to analyze the apoptosis rate.