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Product Detail

Detect changes in intracellular chymotrypsin-like enzyme activity in whole living cells with the FLISP, Fluorescent-Labeled Inhibitors of Serine Proteases, product line of assay kits. This kit utilizes the probe SLCK, which contains a leucine (L) chymotrypsin-targeting amino acid residue linked on the amino terminus with a sulforhodamine 101 (SR) fluorescent reporter tag. The carboxyl end of the L amino acid residue contains a chloromethyl ketone (CMK) reactive 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 probes will interact with the active catalytic sites of chymotrypsin-like proteases, quickly forming covalent bonds with either the reactive site histidine (N-H) (with the CMK-type FLISP probes). Unbound FLISP reagent is easily removed during the wash step, leaving cells with greater quantities of active chymotrypsin-like enzyme activity with a greater fluorescence potential than cells that did not undergo an upregulation of serine protease activity. The red fluorescent SLCK FLISP probe is excited at 590 nm and emit at 620 nm.

FLISP probes are cell permeant and non-cytotoxic at the concentrations suggested in the assay protocol. The SR101-L-CMK FLISP chymotrypsin enzyme detection assay kits can be used in conjunction with your existing apoptosis detection protocols. Each kit includes either 25 tests or 100 tests of SR101-L-CMK reagent, 10X wash buffer, 10X fixative, and Hoechst 33342 dye. SR101-L-CMK stained cells can be analyzed using fluorescence microscopy and fluorescence plate reader evaluation methodology. If a green laser is available on your flow cytometer, this method may be utilized as well.