To determine the percentage of the cell population in each cell cycle phase, we performed ow cytometry with detection of propidium iodide (PI). CAD cells were plated in T75 asks and incubated with different conditions and time as indicated. Next, cells were harvested and washed twice with cold 1X PBS and xed in 70% ethanol at -20°C overnight. On the day of measurement, the pellets were collected by centrifugation and resuspended in 500 µL of PI/RNase A staining buffer (catalog number F10797, Invitrogen, USA). Cells were subsequently incubated in the dark for 15 minutes at room temperature. We then quanti ed the percentage of cells in each gate of cell cycle, based on the DNA content inside the cells, via the BD FACSCanto II ow cytometer (Amersham Biosciences, USA).
Pi rnase a staining buffer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PI/RNase A staining buffer is a laboratory reagent designed for the preparation and staining of samples for flow cytometric analysis. It contains propidium iodide (PI) and RNase A, which are used to stain cellular DNA and remove RNA, respectively. This buffer is commonly used in cell cycle analysis and apoptosis studies.
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Lab products found in correlation
2 protocols using pi rnase a staining buffer
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Cell Cycle Phase Analysis by Flow Cytometry
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Cell Cycle Phase Analysis by Flow Cytometry
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To determine the percentage of the cell population in each cell cycle phase, we performed ow cytometry with detection of propidium iodide (PI). CAD cells were plated in T75 asks and incubated with different conditions and time as indicated. Next, cells were harvested and washed twice with cold 1X PBS and xed in 70% ethanol at -20°C overnight. On the day of measurement, the pellets were collected by centrifugation and resuspended in 500 µL of PI/RNase A staining buffer (catalog number F10797, Invitrogen, USA). Cells were subsequently incubated in the dark for 15 minutes at room temperature. We then quanti ed the percentage of cells in each gate of cell cycle, based on the DNA content inside the cells, via the BD FACSCanto II ow cytometer (Amersham Biosciences, USA).
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