Apoptosis Quantification using Annexin V

Apoptosis was analyzed using the Annexin V-Allophycocyanin/7-Amino Actinomycin D apoptosis Detection Kit (KeyGEN). Briefly, cells in the logarithmic phase were digested with trypsin without EDTA, pelleted at 1200 rpm for 4 minutes, and washed twice with cold PBS. Then, 100 μL Binding Buffer was used to suspend the cells. Subsequently, 5 μL of the 7-Amino Actinomycin D and Annexin V-Allophycocyanin was added and mixed. The cells were incubated at room temperature in the dark for 10 minutes. The cells were then added with 400 µL binding buffer, mixed evenly, and analyzed on ACEA NovoCyte3130 within 1 hour.