Cells were plated in six-well plates, followed by incubation with serum-free medium for 48 hours for cell cycle synchronization, and then recovered serum. Cells were collected at 2-time points, 0 and 24 hours, after serum recovery. The attached cells were harvested and washed with 1 ml PBS, then centrifuged at 1000 rpm for 10 min and discarded the supernatant. Fixed cells with 1 ml pre-cooled 75% ethanol, vortexed, and kept cells at 4 °C overnight. Then centrifuged cells at 1000 rpm for 10 min and discard the supernatant. Stained cells with cell cycle detection kit (BD Pharmingen, #554656) after washed with PBS, resuspend the cells in 0.5 ml PT/RNase staining solution (BD Pharmingen, #550825), 1 × 106 cells per tube, and incubated at room temperature in the dark for 15 min, then detect with NovoCyte Flow Cytometer (ACEA Biosciences).
Pt rnase staining solution
Manufactured by BD
PT/RNase staining solution is a laboratory reagent used to stain cell nuclei and detect the presence of RNA. It contains propidium iodide, a fluorescent dye that binds to nucleic acids, and RNase, an enzyme that degrades RNA. The solution is used in various cell biology and molecular biology applications that require the visualization and quantification of cellular DNA and RNA content.
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Lab products found in correlation
2 protocols using pt rnase staining solution
1
Cell Cycle Synchronization and Analysis
2
Cell Cycle Analysis by Flow Cytometry
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Cells were plated in six-well plates, followed by incubation with serum-free medium for 48 hours for cell cycle synchronization, and then recovered serum. Cells were collected at 2-time points, 0 and 24 hours, after serum recovery. The attached cells were harvested and washed with 1 ml PBS, then centrifuged at 1000 rpm for 10 min and discarded the supernatant. Fixed cells with 1 ml pre-cooled 75% ethanol, vortexed, and kept cells at 4°C overnight. Then centrifuged cells at 1000 rpm for 10 min and discard the supernatant. Stained cells with cell cycle detection kit (BD Pharmingen, #554656) after washed with PBS, resuspend the cells in 0.5 ml PT/RNase staining solution (BD Pharmingen, #550825), 1 × 10 6 cells per tube, and incubated at room temperature in the dark for 15 min, then detect with NovoCyte Flow Cytometer (ACEA Biosciences).
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