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.
Because of their supporting role in the apoptotic process, serine protease activity will be greater in apoptotic cell populations compared to healthy cells of the same type. Quantitate intracellular chymotrypsin-like serine protease activity in vitro with ICT’s FLISP kits. If there is an active chymotrypsin-like enzyme inside the cell, it will covalently bind with the FLISP inhibitor, in this case FAM-Phe-CMK (FFCK), and retain the green fluorescent signal inside the cell. Cells containing lower concentrations of chymotrypsin-like enzyme activity will retain a lower level of fluorescence compared to cells containing higher concentrations of this effector enzyme component.
FLISP may be used in combination with FLICA to discriminate serine protease activity from caspase activity in the same cell. Analyze results using a flow cytometer, fluorescence microscope, or fluorescence plate reader.
FFCK
Chymotrypsin-like enzymes
488 nm / 520 nm
Flow cytometry, Fluorescence microscope, fluorescence plate reader
Cell culture, tissue
FLISP at ≤ -20°C, other components at 2-8°C
Domestic: Overnight Delivery; International: Priority Shipping
United States
- 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.
Kit 945: 25 Tests
FAM-Phe-CMK, 1 vial, #6146
Hoechst Stain, 1 mL, #639
Propidium Iodide, 1 mL, #638
10X Cellular Wash Buffer, 15 mL, #6164
Fixative, 6 mL, #636
Kit Manual
Kit 946: 100 Tests
FAM-Phe-CMK, 4 vials, #6146
Hoechst Stain, 1 mL, #639
Propidium Iodide, 1 mL, #638
10X Cellular Wash Buffer, 60 mL, #6165
Fixative, 6 mL, #636
Kit Manual