Quantitate intracellular chymotrypsin-like serine protease activity in vitro with ICT’s FLISP kits. Chymotrypsin-targeting FLISP probes interact with active catalytic sites of chymotrypsin-like proteases, so cells with greater quantities of active chymotrypsin-like enzyme activity with a greater fluorescence potential than those that are not upregulated. Analyze your samples using a flow cytometer, fluorescence microscope, or fluorescence plate reader.
- Prepare samples and controls
- Dilute cellular wash buffer 1:10 with diH2O
- Reconstitute FLISP with 50 µL DMSO
- Dilute FLISP 1:5 by adding 200 µL PBS
- Add 10 µL FLISP to each sample (~500 µL aliquot of cultured cells)
- Incubate ~ 1 hour.
- Remove media and wash cells: add wash buffer and spin cells (twice); or add fresh media and incubate 1 hour
- If desired, label with additional stains, such as Hoechst, PI, or an antibody
- If desired, fix or embed cells
- Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. FAM-FLISP excites at 488 nm and emits at 520 nm.
Product Specific References
|35861835||Yang, S.F., et al. 2022. Chronic Kidney Disease Is Associated With Increased Cardiac Corin Expression But Decreased Proatrial Natriuretic Peptide Conversion Activity. Journal of the American Heart Association, e025208.|