Apoptosis in the siRNA-treated MCF-7 cells was analysed using flow cytometry (FACSAria II, BD Biosciences). The cultured cells were washed twice with PBS and were resuspended in binding buffer at a concentration of 1 × 106 cells/ml. The cell suspensions (1 × 105 cells/100 μl) were transferred into 5 ml culture tubes, and 5 μl of annexin V-phycoerythrin (eBioscience) and 5 μl of 7-amino-actinomycin (eBioscience) were then added. The cells were gently vortexed and incubated at room temperature in the dark for 15 min. Subsequently, another 400 μl of aliquot of binding buffer were added. Flow cytometry was performed within 4 h of staining.
Annexin 5 phycoerythrin
Manufactured by Thermo Fisher Scientific
Sourced in United States
Annexin V-phycoerythrin is a fluorescent protein conjugate used in flow cytometry analysis to detect and quantify apoptotic cells. It binds to phosphatidylserine exposed on the surface of cells undergoing apoptosis.
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3 protocols using annexin 5 phycoerythrin
1
Apoptosis Analysis in MCF-7 Cells
2
Apoptosis Analysis in MCF-7 Cells
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Apoptosis in the siRNA-treated MCF-7 cells was analyzed using flow cytometry (FACSAria II, BD Biosciences). The cultured cells were washed twice with PBS and were resuspended in binding buffer at a concentration of 1 × 106 cells/mL. The cell suspensions (1 × 105 cells/100 µL) were transferred into 5-mL culture tubes, and 5 µL of annexin V-phycoerythrin (eBioscience) and 5 µL of 7-amino-actinomycin (eBioscience) were then added. The cells were gently vortexed and incubated at room temperature in the dark for 15 min. Subsequently, another 400-µL aliquot of binding buffer was added. Flow cytometry was performed within 4 h of staining.
3
Apoptosis Assay for Anti-PADI2 siRNA
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MNK-45 cells or Bel-7402 cells with the treatment of anti-PADI2 siRNA were washed twice with PBS and were resuspended in binding buffer at a particular concentration. The cell suspensions at a concentration of 1×105 cells/100 μL were transferred into 5 mL culture tubes, and 5 μL of annexin V-phycoerythrin (eBioscience, San Diego, CA, USA) and 5 μL of 7-amino-actinomycin (eBioscience) were added to the tube. The mixture was placed at room temperature in the dark for 15 min. Later, another 400 μL aliquot of binding buffer was added. The apoptosis ratio was examined by flow cytometry (FACSAria II; BD Biosciences, Franklin Lakes, NJ, USA).
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