Fitc conjugated annexin 5 pi assay kit

Apoptosis was determined by using the FITC-conjugated Annexin-V/PI assay kit (Roche, Basel, Switzerland) and flow cytometry following the manufacturer's instructions. Annexin-V-positive cells were classified as early apoptotic cells, and Annexin V and PI double-stained cells were classified as late apoptotic or necrotic cells.

Apoptotic status was determined by FITC-conjugated Annexin-V/PI assay kit (Roche) using flow cytometry following the manufacturer’s instructions. Briefly, 2 × 10⁵ cells were seeded in 6 well flat bottom plates for 24 h and exposed to drugs for 16 h. Dead cells were collected and the remaining cells were rinsed with PBS and detached using trypsin. Detached cells were resuspended in 100 μl binding buffer containing FITC-conjugated Annexin-V (10 mg/mL)/PI (50 mg/ml) and incubated at RT for 15 min. The cells were diluted in 400 μl of PBS and analyzed by a FACScan flow cytometry (Becton Dickinson, Franklin Lakes, NJ USA). Apoptosis and necrosis were evaluated using FL2 (PI) vs FL1 (Annexin V) plots. The cells stained with Annexin V only were classified as early apoptosis and the Annexin V and PI double-stained cells were classified as late apoptosis or necrosis.