Cell preparation was the same with that of the cell proliferation assay. After the treatment of MK2206, cells in the presence or absence of MK2206 were then cultured for another 24 h. The cell cycle was detected by the standard propidium iodide method. Briefly, the treated cells were first trypsinized then fixed with 70% ethanol at 4°C overnight. After being washed twice by PBS, cells were then treated with 100 units/ml RNase A at 37°C for 30 min, followed by staining with 50 mg/ml propidium iodide at 4°C for 30 min with protection from light. The cell cycle proportion was determined by flow cytometric analysis (FACSCanto II system, BD, Franklin Lakes, NJ, USA). The data was analyzed by the software of ModFit LT for Windows Version 3.2, Verity Software House, USA.
Modfit lt for windows version 3
Manufactured by Verity Software House
Sourced in United States
ModFit LT for Windows Version 3.2 is a software application used for the analysis of flow cytometry data. It provides tools for cell cycle analysis, DNA content analysis, and other flow cytometry data processing functions.
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Lab products found in correlation
2 protocols using modfit lt for windows version 3
1
Cell Cycle Analysis by Flow Cytometry
2
Cell Cycle Analysis via Flow Cytometry
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The cells in logarithmic phase were seeded into a culture flask at a density of 1×105/ml for 24 h at 37°C with 5% CO2. The medium was then replaced with medium without FBS to culture for another 24 h at 37°C with 5% CO2. The cells were collected by centrifugation 300 × g for 3 min at room temperature and washed twice with pre-cooled PBS. The 70% pre-cooled ethanol was added to fix at 4°C overnight. The cells were reserved at 4°C and subjected to propidium iodide staining and then the cell cycle was detected using BD flow cytometry (BD Biosciences, San Jose, CA, USA), and data was analyzed using ModFit LT for Windows Version 3.2 (Verity Software House, Topsham, ME, USA).
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