The cells (about 1 × 105 cells) were harvested, cleaned once with phosphate buffer saline, and re-suspended with 500 µL of 1× binding buffer (BioVision, Milpitas, CA, USA). 5 µL of Annexin V-FITC staining solution and 10 µL of PI staining solution (BD Biosciences, San Diego, CA, USA) were supplemented into each sample, and then the cells were incubated in the dark for 30 min. Then the apoptosis was monitored by a flow cytometer (FACScan; BD Biosciences, San Jose, CA, USA).
Annexin 5 fitc staining solution
Manufactured by BD
Sourced in United States
Annexin V-FITC staining solution is a laboratory reagent used for the detection of apoptosis. It binds to phosphatidylserine, which is exposed on the surface of apoptotic cells. The FITC (fluorescein isothiocyanate) label allows for the visualization of Annexin V binding, enabling the identification of cells undergoing programmed cell death.
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3 protocols using annexin 5 fitc staining solution
1
Annexin V-FITC/PI Apoptosis Assay
2
Apoptosis Quantification in Transfected Cells
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The transfected cells were inoculated into a 6-well plate (1 × 105 cells/well) and cultured overnight, and then the cells were trypsinized with trypsin, washed with Phosphate Buffer Saline (PBS), harvested, and re-suspended in 300 μL of binding solution. Then the cells were mixed with and incubated with 10 μL of Annexin V-FITC staining solution (BD Bioscience, San Diego, CA, USA) for 15 min in the darkness at ambient temperature. Then, 5 μL of Propidium Iodide (PI) staining solution (BD Bioscience, San Diego, CA, USA) was added for staining the cells for 5 min in darkness. Finally, the cells were washed with PBS, and the apoptosis of the transfected cells was detected by a flow cytometer.
3
Annexin V-FITC/PI Cell Apoptosis Assay
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After treatment with PYM at 0.1, 1, 10, and 100 µg/mL for 24 h, which was stopped by FBS addition, cells in the treatment and control groups were collected, washed twice with PBS (0.01 mM), and then transferred to Eppendorf tubes.
Annexin V-FITC (200 µL) was added into each tube, and then 5 µL Annexin V-FITC staining solution and 5 µL of propidium iodide (PI) solution were added into the treatment and control groups (from BD Biosciences, San Diego, CA). After staining in the dark for 15 min and on ice for 10 min, all tubes were analyzed by flow cytometry (BD Biosciences).
Annexin V-FITC (200 µL) was added into each tube, and then 5 µL Annexin V-FITC staining solution and 5 µL of propidium iodide (PI) solution were added into the treatment and control groups (from BD Biosciences, San Diego, CA). After staining in the dark for 15 min and on ice for 10 min, all tubes were analyzed by flow cytometry (BD Biosciences).
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