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Flow cytometry to detect apoptosis

WZ Wenjuan Zha
XL Xiaomin Li
XT Xiaowei Tie
YX Yao Xing
HL Hao Li
FG Fei Gao
TY Ting Ye
WD Wangqi Du
RC Rui Chen
YL Yangchen Liu
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An Annexin V-FITC Apoptosis Detection Kit (KeyGen Biotech, Nanjing, China, KGA108) was used to detect cell apoptosis. Cells were inoculated on a six-well plate and washed twice with pre-cooled PBS at 24 h after transfection. The cells were digested with trypsin without EDTA, centrifuged at 300×g for 5 min at 4 °C, washed once with pre-cooled PBS, and centrifuged at 4 °C at 300×g for 5 min. Then, 250 µl of binding buffer was added to the cells to resuspend them, and 100 µl of cells was added to a flow tube. A PI solution (5 µl) and Annexin V-Alexa Fluor (10 µl) were mixed and reacted with the cells at room temperature for 15 min, after which flow cytometry analysis was performed.

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