Hi, I am planning to use the MitoTracker Green FM dye for HEK293 cells, have you used HEK293 for such experiment? What concentration normally one uses for HEK293 cells and what should be the incubation time?
Apart from that I am concerned about using DMEM +10% FBS medium for labeling the cell with dye, as manual says proteins present in Serum may oxidize dye. So how do one go about it? As I have an experience of cells coming out very easily if kept in PBS for longer time which is advised in manual.
use 200nM concentration of mitotracker. Incubate your cells for half an hour In media contaning the dye.
Thanks Ankit, Do you face any problem with Phenol Red containing media for fluorescence microscopy, as phenol red may give some back ground?
No, phenol red doesn't give you any background. Only if you use high concentration of dye that will cause background. But don't worry, if you see too much background you can discard the medium and add PBS.
Joel, For any imaging experiment, avoid using phenol-red containing media.Especially if you are looking at ER / Mitochondria. You can get phenol-red free media from HiMedia (very cheap) and works pretty well.
Or go to Diblab.
Thanks Abira di :) Bhavik only advised me to use phenol red free medium. :)
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