Method for measuring target cell killing - using flow cytometry

To do immunology, especially oncology experiments, it is inevitable to test the killing of tumor cells by lymphocytes or other cells. The most commonly used method in foreign countries is the Cr51 radiation experiment, but this method can not be done in many places in China, and the method of MTT It is too old and not accurate, so the alternative method uses flow cytometry, which is convenient and accurate.

The specific reference is as follows:
1. Count the target cells and add CFSE to achieve a final CFSE concentration of 2 uM for 37 minutes and 30 minutes.
2, after taking out, 1000rpm, 5min, you can see the cells stained yellow, washed three times with PBS
3, mixed with effector cells, for 4-6 hours.
4, the cell mixture is digested and removed, washed twice, PI working solution 5ul, protected from light for 30 minutes, upflow
5, CFSE, PI double positive cells are killed target cells, divided by the total number of target cells, that is, the kill rate.

Precautions:
1, CFSE and PI concentration must pay attention to, Ning low is not high, personal feeling, if the elevation, it would be better to put one night at 4 degrees
2, must be set to the whole negative, CFSE single standard, PI single standard, CFSE + PI double standard and synchronized target cell reference.
3, the action time can not be too long, not more than 8 hours.

Biological reagents you might be interested in:

Tumor/apoptosis research related antibody human, large mouse, etc. Elisa detection kit growth factor chemokine protease neurotrophin