Magic RedFluorescent Cathepsin L Assay Kit

1. Prepare samples and controls, 2. Dilute cellular wash buffer 1:10 with diH2O, 3. Reconstitute FLISP with 50 µL DMSO, 4. Dilute FLISP 1:5 by adding 200 µL PBS, 5. Add 10 µL FLISP to each sample (~500 µL aliquot of cultured cells), 6. Incubate ~ 1 hour., 7. Remove media and wash cells: add wash buffer and spin cells (twice); or add fresh media and incubate 1 hour, 8. If desired, label with additional stains, such as Hoechst, PI, or an antibody, 9. If desired, fix or embed cells, 10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLISP excites at 488 nm and emits at 520 nm.

Question: When using a fluorescence plate reader staining for adherent cell, do I have to detach the cell by trypsin treatment? In that case, Is there anything to be aware of?, , Answer: If you are using adherent cells, you do not necessarily need to detach them. If your plate reader is capable of reading from the bottom, you can grow your adherent culture in a tissue culture plate with black walls and a clear bottom.