Annexin 5 apc pi apoptosis detection kit

Cell apoptosis was evaluated by flow cytometry using an Annexin V-APC/PI Apoptosis Detection Kit (KeyGen BioTECH, Nanjing, China). Cells with different treatment were incubated in a 6-well plate for 48 hours. All cells were trypsinized and the resuspended cells were washed twice with PBS. Cells were then resuspended in 500 µL binding buffer, and 5 μL Annexin V-APC and 5 μL propidium iodide (PI) were added into cell suspension at room temperature for 15 min in the dark before detection.

Annexin V-APC/PI Apoptosis Detection Kit (KeyGEN, China) was employed to make flow cytometry analysis for apoptosis. Cells were harvested at 48 hours after transfection, followed by resuspending in 500ul binding buffer. Then, away from light, 5ul Annexin V-APC and 5ul propidium iodide (PI) were put at room temperature for 15 min. Cell apoptosis was determined using CytoFLEX Flow Cytometer (Beckman, USA) within a 1 h period and data was analyzed using FlowJo software.

The cell apoptosis assay for stably expressed THP-1 cells was conducted using the Annexin V-APC/PI Apoptosis Detection Kit (KeyGEN, Jiangsu, China) following the manufacturer’s instruction. In brief, the cells were collected and washed with 1× phosphate-buffered saline (PBS, Solarbio, Beijing, China), added with 3% FBS three times, and then incubated with Annexin V-APC/PI for 15 min. In the CytoFLEX flow cytometer (Beckman Coulter, CA, United States), the FL3 channel was used for detection of PI and FL4 channel was used to detect Annexin V-APC. The results were analyzed by CytExpert software (Beckman Coulter, CA, United States). The experiments were performed at least in triplicate.