OS MG63 cells were seeded into 96-well culture dishes at 106 cells/ml and 2-ME added to 0, 10, 20 or 40 µM (6 wells/group). After 2 days of culture, trypsin was added and the cell suspensions centrifuged (1,000 × g for 5 min at room temperature). The cells were collected and incubated with RNase A for 30 min, propidium iodide (PI) reagent (BD Biosciences) was then added and the cells incubated in the dark for a further 30 min. Finally, the cells were analyzed by flow cytometry using a FACSCanto II (BD Biosciences), and data were analyzed using Cell Quest Pro software version 5.1 (BD Biosciences).
Pi reagent
Manufactured by BD
Sourced in United States
PI reagent is a laboratory product used for cell analysis. It is a fluorescent dye that binds to DNA and is commonly used to stain and identify cells. The core function of PI reagent is to enable the detection and quantification of cellular DNA content through flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
3 protocols using pi reagent
1
Cell Cycle Analysis of MG63 Cells
2
Apoptosis Evaluation by Flow Cytometry
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Cells were collected 24 h after transfection, washed twice with pre-chilled 1 × PBS, and resuspended to a density of approximately 1 × 106 cells/ml. 100 µL of the cell suspension was transferred to a 1.5 ml EP tube, and 5 µL of FITC Annexin V, and 5 µL of PI reagent (556547; BD Biosciences, San Diego, CA, USA) were added and mixed well. Incubate for 15 min at room temperature in the dark. Flow cytometry was then performed.
3
Apoptosis and Cell Cycle Analysis
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HFLS-RA cells were transfected and harvested after 48 h, and processed as per the PE Annexin V Apoptosis Detection Kit (BD, USA). Briefly, 5 μl of the PE reagent and 5 μl of 7-AAD reagent were added into the cell suspensions and incubated for 15 min at room temperature. Finally, we analyzed the cells in different phases using the FACSAria flow cytometer (BD).
HFLS-RA cells were harvested 48 h after transfection and fixed with 75% ethanol at −20°C for at least 2 h. Subsequently, cells were resuspended in the PI reagent (BD). Finally, we analyzed the cells in different phases using the FACSAria flow cytometer (BD).
HFLS-RA cells were harvested 48 h after transfection and fixed with 75% ethanol at −20°C for at least 2 h. Subsequently, cells were resuspended in the PI reagent (BD). Finally, we analyzed the cells in different phases using the FACSAria flow cytometer (BD).
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