I did the cell penetrating test for the novel peptide labelled with FITC. I incubated cell in serum free medium (DMEM) for 4 hrs. Afterward I washed with PBS and incubated cell with fetal bovine containing serum for 2 hrs.
My results were unexpected - I got the fluorescence at 488 wavelength in control plate.
Does fetal bovine serum have fluorescence at 488?
I expect it does. I have worked with mouse and with human serum and both produced a very high background with 488 nm excitation. I was using FLCS method and the differences in fluorescence lifetimes between the serum and the fluorophore to observe the signal from the fluorophore (which was actually contributing to the total intensity less than the serum)
Thank you ...
For your help...I think also. i repeated the experiment and found that it has some background..so it is mostly because of serum fluorescence...
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