Flow Cytometric Analysis of Cell Cycle

Flow Cytometric Analysis of Cell Cycle

Contributor: Suprya Jayadev Date: July 13, 1994 Fixation 1) Collect 2 X 106 cells. 2) Pellet cells by spinning at 1,000 rpm, 4°C for 5 minutes. 3) Resuspend cell pellet in 1 ml of cold PBS. 4) Fix cells by adding 4 ml of -20°C absolute ethanol. 5) Store cells at -20°C in this fixation buffer until ready for analysis. (No more than 2 weeks) Staining 6) Centrifuge (as above) fixed cells and resuspend pellet in 1 ml of PBS. 7) Add 100 µl of 200 µg/ml DNase-free, RNaseA and incubate at 37°C for 30 minutes. 8) Add 100 µl of 1 mg/ml propidium iodide (light sensitive) and incubate at room temperature for 5-10 minutes. 9) Place samples in 12 X 75 Falcon tubes and read on Becton Dickinson FACStarPLUS.

ReferenceExperimental Cell Research 207 ,142-151.