• Directly detect activities using cell-permeant reagents
• Labeled inhibitors ensure no cross-reaction with inactive enzyme
• No antibodies
• No lysing, fixing, or permeabilization
• Red or fluorophores available to detect apoptosis via active caspases

ICT's FLISP Kits utilize cell-permeable, fluorescently labeled physositigmine -- a cholinesterase inhibitor, linked to a green fluorophore (Ph-Fl) -- to detect and localize active cholinesterase enzyme in whole living cells. Because Ph-Fl reagents are cell-permanent, you don't need to lyse the cells or permeabilize the membranes, nor do you have to worry about cross-reaction with inactive or pro forms of the enzyme. Only cells with the active cholinesterase will fluoresce.

When used in combination with FLICA detection kits, which detect active caspases using a red-florescent inhibitor analog, both cholinesterase and caspase activities can be simultaneously monitored in the same cells. Cells with active caspases fluoresce red and those with active cholinesterase fluoresce green -- providing two complementary assays that enable you to clearly identify apoptotic cells (see figure). Positive cells can be detected with a fluorescence microscope, a fluorescence plate reader, or a flow cytometer. You can analyze individual cells or rapidly probe large numbers of cells with high accuracy and sensitivity.

A	B
Figure A, B: Varying expressions of cholinesterase and caspase activity in apoptosis-induced Jurkat cells stained with Ph-Fl and SR-VAD-FMK. Cell 1 has active cholinesterase (green) but has no caspase activity (not stained red). Cells 2 and 3 reveal cholinesterase and caspase activity at different intensities (green and red).

• Cholinesterase Detection Kit

Try the Cholinesterase kits with ICT's SR FLICA™ Apoptosis Kits (red fluorescent inhibitors) for dual-staining studies.