Annexin 5 fitc pi apoptosis detection kit

Cell apoptosis was detected by Annexin V-FITC/PI Apoptosis Detection Kit (E606336, Sangon Biotech Co., Ltd., China). Cells were rinsed with PBS and suspended in 1 × binding buffer (195 μL) at a density of 2 × 10⁵ cells/mL. Next, 5 μL Annexin V-FITC was added and mixed well in a dark room, followed by culture for 15 min at room temperature. Then, cells were washed by 1 × binding buffer (200 μL) and centrifuged for 5 min at 1000 rpm. As the supernatant was removed, cells were resuspended in 1 × binding buffer (190 μL) with 10 μL PI. A CytoFLEX S flow cytometer (Beckman Coulter, Inc., Brea, CA, USA) was adopted for cell apoptosis analysis.

The Annexin V-FITC/PI Apoptosis Detection Kit (E606336, Sangon Biotech Co., Ltd., China) was applied for cell apoptosis assessment. Briefly, 4 × binding buffer was diluted into 1 × binding buffer, in which cells were suspended and adjusted to a density of 4 × 10⁵ cells/ml. Next, 5 μl Annexin V-FITC was added into 195 μl cell suspension and mixed well in a dark room, followed with incubation for 10 min at room temperature. Then, cells were rinsed using 1 × binding buffer (200 μl) and centrifuged for 5 min at 1,000 ×g. Supernatant removed, cells were resuspended in 190 μl binding buffer (1 ×) and 10 μl PI added. Cell apoptosis was analyzed using a flow cytometer (Multiskan FC, Thermo Fisher Scientific).

Cyclo-(DP)₃ was purchased from Shanghai Qiangyao Biological Technology (Shanghai, China). N, N-dimethylformamide (DMF), triethylamine (TEA), and CH3I were obtained from TCI (Shanghai) Development Co., Ltd. (Shanghai, China). Lipofectamine 2000 was provided by Thermo Fisher Scientific (Shanghai, China). Dimethylsulfoxide, hexamethylenediamine, and 2,2′-diamino-N-methyldiethylamine were purchased from Solarbio Life Sciences (Beijing, China). Survivin-siRNA, FAM-siRNA, and scrambled siRNA were purchased from GenePharma (Shanghai, China). Primers were purchased from Comate Bioscience (Changchun China). MTT was obtained from Genview (Beijing, China). DAPI, TRIzol reagent, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel preparation kits were acquired from Solarbio Life Sciences (Beijing, China). All-In-One RT MasterMix kits were purchased from TransGen Biotech (Beijing, China). The UNIQ-10 Column Total RNA Extraction Kit, Go Taq qPCR Master Mix Kit, and Annexin V-FITC/PI Apoptosis Detection Kit were purchased from Sangon Biotech (Shanghai,China). Anti-survivin antibody A5719 was obtained from Bimake (Houston, TX, USA). GAPDH (rabbit, AP0066) was obtained from Bioworld (Minneapolis, MN, USA).