Briefly, 1×105 cells were seeded in a well of a 6-well plate and maintained in medium containing 10% FBS. After 48 h, the cells were harvested and then resuspended with cold PBS. For cell cycle analysis, cells were fixed in 70% ethanol overnight at 4 °C. Fixed cells were washed with PBS, incubated with 50 µg/ml propidium iodide (BD Pharmingen, USA) and 100 µg/ml RNaseA in the dark for 30 min. Cell cycle phase distribution was assessed using a FACScan flow cytometer (BD Biosciences, USA), and data were analyzed using Becton-Dickinson Cell Quest software. For apoptosis analysis, the cells were resuspended in binding buffer and incubated with Annexin V-APC/PI (eBioscience, USA) at room temperature for 15 min. After incubation, the percentage of apoptotic cells was measured with a FACSCalibur flow cytometer (BD Biosciences, USA).
Annexin 5 apc pi
Manufactured by Thermo Fisher Scientific
Sourced in United States
Annexin V-APC/PI is a lab equipment product that is used to detect and quantify apoptosis, a form of programmed cell death, in cells. It uses a combination of Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a fluorescent dye that binds to DNA, to differentiate between viable, apoptotic, and necrotic cells.
Automatically generated - may contain errors
Lab products found in correlation
Propidium iodide, by BD (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Facscan flow cytometer, by BD (1 mentions)
Annexin 5 fitc pi, by Thermo Fisher Scientific (1 mentions)
Hcd34 apc, by Thermo Fisher Scientific (1 mentions)
Facs fortessa, by BD (1 mentions)
Facs c6, by BD (1 mentions)
3 protocols using annexin 5 apc pi
1
Cell Cycle and Apoptosis Analysis
2
Apoptosis Assessment in Leukemia Cells
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To assess apoptosis, CD34+CD38− KG1α or Kasumi cells were cultured as described above for 24, 48, or 72 h with or without chidamide, Ara-C, DNR, or IDA, then double labeled with Annexin V-APC/PI (eBioscience, San Diego, CA, USA) for 15 min at room temperature in the dark according to the manufacturer’s instructions. Primary samples were stained with hCD34-APC (eBioscience, USA) and Annexin V-FITC/PI to assess the apoptosis of CD34+ primary cells induced by chidamide or IDA alone or the two drugs in combination. The stained cells were analyzed by flow cytometry (FACS Fortessa, BD Biosciences). Apoptotic cells were defined as Annexin V positive.
3
Apoptosis Quantification by Annexin V-APC/PI
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Apoptosis was measured using Annexin V-APC/PI (Ebioscience, San Diego, USA) dual staining by flow cytometry. Briefly, cells (2×105/well) were seeded into 24-well plates and exposed to DS at different doses (0.025, 0.05, 0.1, 0.2 μM) either with or without Cu (0.5 μM) for 24 hrs. Cells were harvested, washed twice with iced PBS, and double labeled with Annexin V-APC/PI for 30 minutes at 4°C in the dark. Cells were then analyzed by flow cytometry using FACS C6 (BD, Oxford, UK).
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