After transfection, cell cycle analysis was performed using a cell cycle analysis kit (Lianke, Shanghai, P.R. China). Briefly, cells (4 × 105 per well) were seeded onto six-well plates and starved in FBS-free medium for 12 h. Cells were then washed with PBS, digested by trypsin, and collected by centrifugation at 1,000 rpm for 5 min. After centrifugation, DNA staining was performed with 10 mg PI/ml PBS and 2.5 mg of Ag DNase-free RNase/ml PBS for 30 min. Cell cycle profiles were generated with Modifit software (BD Biosciences, Franklin Lakes, NJ, USA).
Cell cycle analysis kit
Manufactured by Liankebio
Sourced in China
The cell cycle analysis kit is a laboratory tool used to study the different stages of the cell cycle. It provides a standardized method for measuring the distribution of cells in different phases of the cell cycle, including G0/G1, S, and G2/M phases.
Automatically generated - may contain errors
6 protocols using cell cycle analysis kit
1
Cell Cycle Analysis by Flow Cytometry
2
Cell Cycle Analysis by Flow Cytometry
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Cell cycle analysis was performed by Cell Cycle Analysis Kit (Lianke, P.R. China). Cells (4 × 105 per well) were seeded into six-well plates for 24 h and starved in FBS-free medium for 12 h. Cells were then fixed with 70% cold ethanol for 2 h. DNA staining was carried with 10 mg of propidium iodide/ml PBS and 2.5 mg of Ag DNase-free RNase (Roche Diagnostics, Basel, Switzerland)/ml PBS for at least 30 min. Flow cytometry was performed by Coulter EPICS XL flow cytometer (Beckman Coulter, Inc., Fullerton, CA, USA).
3
Cell Cycle Analysis Protocol
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Cell cycle analysis was performed using cell cycle analysis kit (Lianke, Shanghai, China). Briefly, cells were digested with trypsin and washed with phosphate‐buffered saline (PBS). Cells (2 × 105) cells were suspended with 50 μL binding buffer and then treated with cell cycle agents for 48 hours. Then, the adherent cells were washed and trypsinized, and centrifuged at 12 000 g for 5 minutes. After centrifugation, DNA staining was performed with 10 mg PI/mL PBS and 2.5 mg Ag DNase‐free RNase/mL PBS for 30 minutes. Cell cycle profiles were generated with Modifit software (BD Biosciences).
4
Cell Cycle Analysis by Flow Cytometry
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Cell cycle analysis was operated by Cell Cycle Analysis Kit (Lianke, P.R. China). Cells (4 × 105 per well) were seeded into six-well plates for 24 h and starved in FBS-free medium for 12 h. The transfected cells were then harvested and fixed with 500 μl of 70% cold ethanol for 2 h. After centrifugation at 2,000 rpm for 10 min, DNA staining was carried out with 1 ml of propidium iodide (PI) at room temperature for 30 min. Cell cycle distribution was analyzed by measuring DNA content using flow cytometry, and profiles were generated using flow cytometry with Modifit software (BD Biosciences, San Jose, CA, USA).
5
Cell Cycle Analysis of Transfected Cells
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After transfection, cell cycle analysis was performed using a cell cycle analysis kit (Lianke, Shanghai, P.R. China). Briefly, cells (4 × 105 per well) were seeded into six-well plates and starved in FBS-free medium for 12 h. Cells were then washed with PBS, digested by trypsin, and collected by centrifugation at 1,000 rpm for 5 min. The cells (106 cells per sample) were fixed in 4 ml of 75% cold ethanol at −20°C overnight. After centrifugation, DNA staining was performed with 10 mg propidium iodide (PI)/ml PBS and 2.5 mg Ag DNase-free RNase/ml PBS for 30 min. Cell cycle profiles were generated with Modifit software (BD Biosciences, Franklin Lake, NJ, USA).
6
Cell Cycle and Apoptosis Analysis
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Cell cycle and apoptosis analysis were performed using the cell cycle analysis kit (Lianke, Shanghai, P.R. China) and Annexin-V/Dead Cell Apoptosis Kit (Invitrogen). Briefly, cells were seeded in six-well plates at a density of 4 × 105 per well. Cells were starved in FBS-free medium for 12 h and then treated with cell cycle and apoptosis agents for 48 h. The adherent cells were washed and trypsinized, and centrifuged at 12,000 × g for 5 min. The collected cells were fixed in 75% cold ethanol at −20°C overnight and resuspended in annexin-binding buffer and 5 μl of 7AAD/annexin V/propidium iodide at room temperature for 30 min, and then incubated at room temperature for 15 min in the dark. Fluorescence was analyzed by flow cytometry.
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