Quantify intracellular active chymotrypsin-like serine protease activity in vitro. The CMK group alkylates the active site histidine residue. Analyze your samples using a flow cytometer, fluorescence microscope, or fluorescence plate reader.
Samples:
Cell culture, tissue
Application Notes:
Application Notes: 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
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