In brief, PC3 and C4-2 cells were harvested after CVB (80 μmol/L) treatment for 12 h in a 6 cm dish, washed twice in cold PBS and fixed overnight with 70% ethanol. The next day, the fixed cells were washed with PBS again and were then stained with 1 ml of PI/RNase A (BD Pharmingen, Franklin Lakes, NJ, USA) at 37°C in a darkroom for 30 min. A flow cytometer (BD Beckman Coulter, USA) was used to determine the cell cycle distribution. Three independent experiments were conducted with three technical replicates each.
Pi rnase a
Manufactured by BD
Sourced in United States
PI/RNase A is a laboratory product used to extract and purify plasmid DNA from bacterial cultures. It contains a buffer solution and RNase A enzyme, which selectively degrades RNA, allowing for the isolation of pure plasmid DNA.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using pi rnase a
1
Cell Cycle Analysis of CVB-Treated PC3 and C4-2 Cells
2
Cell Cycle Analysis by Flow Cytometry
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To determine the cell cycle distribution, about 2×105 cells were collected in a flow tube after drug treatment and fixed in 70% ethyl alcohol at −20°C overnight. Then, the cells were incubated with PI RNase A (BD Biosciences, Franklin, NJ, USA) for 15 mins. The fractions of the cells in the G0/G1, S and G2/M phases were analyzed by flow cytometry.
3
Cell Cycle Analysis by Flow Cytometry
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The cells were treated with chidamide (1 µM) for 48 h. Subsequently, the cells were collected, washed with PBS and then fixed overnight in 75% ice-cold ethanol at 4°C. The fixed cells were then harvested, stained with propidium iodide (PI)/RNase A (BD Pharmingen; BD Biosciences) and incubated in the dark at room temperature for 30 min after being washed with PBS. The DNA content was analyzed by flow cytometry (FCM) with an LSR2 instrument (BD Biosciences). FlowJo 7.6 software (Tree Star, Inc.) was used for data analysis.
4
Cell Cycle Analysis by Flow Cytometry
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Cells of each group were collected and rinsed twice with PBS. The pre‐chilled ethanol was slowly added to stabilize the cells and stored for later use at −20°C. The cells were resuspended in 500 µL PI/RNase A (550825, BD, USA) staining solution and incubated in the dark for 20 min. The flow cytometry (FACSCalibur, BD, USA) was be used to analyze cell cycle. Flow detection voltage settings was as follows: FSC 319, SSC 324, and PI 429. The experiment was repeated three times.
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