PI/RNase Staining Buffer

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	- Cells were fixed with pre-cold ethanol at 4°C overnight.

Protocol tips
- Cells were fixed with pre-cold ethanol at 4°C overnight.

Publication protocol

At least 1 × 104 transfected U251MG or A172 cells were rinsed with PBS, trypsinized and fixed in 70% pre-cold ethanol at 4°C overnight. Then cells were stained with 500μl PI/RNase Staining Buffer (BD, 550825) at room temperature for 15 min. Cell counts in different cell cycle phase were measured by CyAn ADP 9 colors flow cytometry (Beckman Coulter, USA) and the data were analyzed by Modfit LT 4.1v software. For apoptosis assay, 2 × 105 transfected U251MG or A172 cells were trypsinized, and then co-incubated with Pharmingen™ PE Annexin V(BD, 554656) and 7-Amino-Actinomycin D (7-AAD)(BD, 559925) for 15 min at 4°C in lucifuge condition. After vortex for 15s, the ratio of apoptosis was measured by CyAn ADP 9 colors flow cytometry and the outcome were obtained via Summit V4.3 software.

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