2.7 Flow cytometry determination of caspase-3 protein expression level

2.7 Flow cytometry determination of caspase-3 protein expression level

Preparation of single-cell suspension and FCM detection of caspase-3 protein: Take 0.5 g of tumor tissue, add about 1 ml of 0°C PBS buffer, use ophthalmic surgical scissors to chop the tumor tissue into small pieces, pipette the PBS to rinse it, then filter the solution through a 200-mesh sieve, collect the filtrate in a 10 ml centrifuge tube, centrifuge at 1000 r/min for 5 min, discard the supernatant. Add 5 ml of 0°C PBS, centrifuge at 1000 r/min for 5 min, discard the supernatant. Dilute the precipitate with 0°C PBS and adjust the cell count to more than 10/ml, then load it into a Dorf tube. Follow the FCM operating procedures and the instructions of the rh-Annexin-V-FITC/PI kit to detect the expression of caspase-3 protein.