Matters needing attention in apoptosis detection kit
The attention should be carefully read to ensure that the desired experimental results are prior to use of this kit.
1. The reagent of the screw cap centrifuge tube shall be briefly centrifuged before opening the cover, and the liquid on the inner wall of the cap is thrown to the bottom of the pipe, and the liquid is prevented from falling when the cap is opened.
2. Annexin V-FITC and Propidium iodide are light-sensitive substances and should be protected from light in storage and operation.
3. Propidium iodide (PI) can be absorbed by the skin and has a stimulating effect on the eyes.
4. In the final step of cell washing, discard the supernatant as much as possible in order to avoid the effects of PBS on the experimental results.
It is suggested that the annexin V-FITC staining should be carried out first, and then the PI staining should be added 5 minutes before processing.
6. The whole operation process should be as gentle as possible, do not blow the cells as hard as possible, as far as possible at 4 ℃.
7. Please detect as soon as possible after the reaction, because apoptosis is a dynamic process, and the fluorescence intensity begins to decay an hour after the reaction.
8. The successful detection of apoptosis is affected by the following factors, such as cell type, the density of PS on the cell membrane, the proportion of PS reversal when apoptosis occurs, the method of inducing apoptosis, the reagent used, the time of inducing apoptosis and so on. It is very necessary to optimize these influencing factors for the success of the experiment.