Magic RedFluorescent Cathepsin L Assay Kit

1. Prepare samples and controls, 2. Reconstitute Magic Red by adding 50 µL DMSO., 3. Dilute Magic Red 1:10 by adding 450 µL diH2O., 4. Add 20 µL Magic Red to each sample (~ 500 µL aliquot of cultured cells)., 5. Incubate while protected from light., 6. Watch color start to develop within 15 minutes., 7. If desired, label with additional stains, such as Hoechst, DAPI, Acridine orange, or an antibody., 8. If desired, fix or embed cells., 9. Analyze with a fluorescence microscope or fluorescence plate reader. Magic Red has an optimal excitation at 592 nm and emission at 628 nm., , If working with adherent cells, please see the manual for additional protocols.

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.