Go Back FAM-Spacer-Leu-CMK FLISP Assay Kit--FSLCK

Product Detail

Detect changes in intracellular chymotrypsin-like serine protease enzyme activity in whole living cells with the FLISP, Fluorescent-Labeled Inhibitors of Serine Proteases, product line of assay kits. This assay utilizes a reagent (FSLCK) containing a leucine (L) chymotrypsin-targeting amino acid residue linked on the amino terminus with a carboxyfluorescein (FAM) fluorescent reporter tag. The carboxyl end of the L amino acid residue contains a reactive chloromethyl ketone (CMK) group that targets the catalytic site of serine proteases.

After quickly penetrating the lipid bilayer membranes of the target cell population, the chymotrypsin-targeting FLISP probe FSLCK will interact with the active catalytic sites of chymotrypsin-like proteases, quickly forming covalent bonds with the reactive site histidine (N-H). Unbound FLISP reagent is easily removed during the wash step, resulting in cells with greater quantities of active chymotrypsin-like enzyme activity showing a greater fluorescence potential than cells that did not undergo an upregulation of serine protease activity. Green FAM-FLISP probes like FSLCK are excited at 488 nm and emit in the green wavelength range of 525 nm. Red fluorescence FLISP probes are also available.

FLISP probes are cell permeant and non-cytotoxic at the concentrations suggested in the assay protocol. The FAM-spacer-L-CMK chymotrypsin enzyme detection assay kits can be used in conjunction with existing apoptosis detection protocols. Each FLISP FSLCK kit includes either 25 tests or 100 tests of FAM-spacer-F-CMK reagent, 10X wash buffer, 10X fixative, PI vital stain, and Hoechst 33342 dye. FSFCK-stained cells can be analyzed using flow cytometry, fluorescence microscopy, and fluorescence plate reader evaluation methodology.