Annexin 5 apc propidium iodide apoptosis detection kit

The Annexin V-APC/propidium iodide (PI) apoptosis detection kit (KeyGEN, China) was used to detect apoptosis by flow cytometry. DLBCL cells were cultured in vitro and divided into nine groups, including untreated (blank control), negative control, Astragali Radix group, Glycyrrhizae Radix et Rhizoma group, Ginseng Radix et Rhizoma group, Pseudostellariae Radix group and Codonopsis Radix group. After cultivation for 48 h, cells were washed with PBS, resuspended in 100 µL binding buffer at a concentration of 2 × 10⁶ cells/ml, and anti-Annexin V APC-conjugated antibody and PI were added. The mixtures were incubated for 15 min at room temperature, supplemented with binding buffer to 500 µL and processed by BD Accuri C6 Plus Flow Cytometry (BD Biosciences, United States). Data were analyzed through flow cytometry (BD PharmingenTM, United States).

Cell apoptosis was detected using the Annexin V-APC/propidium iodide (PI) apoptosis detection kit (KeyGen Biotech Co., Ltd., Nanjing, China) according to the manufacturer’s instructions. In brief, we seeded 5 × 10⁵ cells in a 60mm culture dish and collected 5 × 10⁵ cells after 24 h using trypsin, followed by three washes with PBS. We then resuspended the cells in 500 μl of binding buffer, added 5 μl of Annexin V-APC, and added 5 μl of PI. After incubating in the dark for 10 min, flow cytometry (ACEA Bioscience, USA) analysis was performed, with 2–4×10⁴ cells analyzed in each run.