SR101 FLISP SLCK Serine Protease Assay

$185.00$510.00

Quantitate intracellular chymotrypsin-like serine protease activity in vitro with ICT’s FLISP kits. The FLISP inhibitors are labeled with either a green carboxyfluorescein (FAM) or red sulforhodamine 101 (also known as Texas Red™) fluorochrome, contain either the target amino acid leucine or phenylalanine, and a reactive group that targets the catalytic sites of serine proteases. The chloromethyl ketone (CMK) reactive group irreversibly binds to the active site histidine residue present in serine proteases. Cells with greater quantities of chymotrypsin-like enzyme activity will demonstrate a greater fluorescence potential than those exhibiting baseline serine protease activity. Analyze your samples using a flow cytometer, fluorescence microscope, or fluorescence plate reader.

Catalog # Size Quantity Price
955 25 tests $185.00
956 100 tests $510.00
Catalog #: 955, 956 Categories: , ,

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 SR101-Leu-CMK (SLCK), and retain the red 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.

1. Prepare samples and controls
2. Dilute cellular wash buffer 1:10 with diH2O
3. Reconstitute FLISP with 50 µL DMSO
4. Dilute FLISP 1:5 by adding 200 µL PBS
5. Add 10 µL FLISP to each sample (~500 µL aliquot of cultured cells)
6. Incubate ~ 1 hour.
7. Remove media and wash cells: add wash buffer and spin cells (twice); or add fresh media and incubate 1 hour
8. If desired, label with additional stains, such as Hoechst, PI, or an antibody
9. If desired, fix or embed cells
10. Analyze with a fluorescence microscope, fluorescence plate reader, or flow cytometer. SR-FLISP excites at 590 nm and emits at 610 nm.

Suspension cells were treated with an agent to induce serine protease activity, (bottom), or DMSO, a negative control (top), then incubated with ICT’s red serine protease inhibitor probe, SR-FLISP™, washed, and analyzed with a flow cytometer. The experimental reatment induced serine protease activity in 56.3% of the experimental cells (M2, bottom right), whereas the negative control treatment exhibited serine protease activity in only 13.5% of the cell population (M2, top right). This is a ratio of 4:1. (Dr. Brian W. Lee & Sally Hed, ICT).srflispdata

• Reagent name: SLCK
• Target: Chymotrypsin-like enzymes
• Excitation/Emission: 590 nm / 610 nm
• Method of Analysis: Flow cytometry, Fluorescence microscope, fluorescence plate reader
• Types of Samples: Cell culture
• Storage: FLISP at ≤ -20°C, all other kit components at 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping

Kit 955: 25 tests
• SR101-Leu-CMK, 1 vial, #6152
• Hoechst Stain, 1 mL, #639
• 10X FLISP Wash Buffer, 15 mL, #6164
• Fixative, 6 mL, #636
• Kit Manual

Kit 956: 100 tests
• SR101-Leu-CMK, 4 vials, #6152
• Hoechst Stain, 1 mL, #639
• 10X FLISP Wash Buffer, 60 mL, #6165
• Fixative, 6 mL, #636
• Kit Manual