Article No
260-963
Application | Activity Assay |
Article No | 260-963 |
Country Availability | SE, FI, DK, NO, IS, EE, LV, LT |
Components | Kit 963: 25 tests • FAM-Spacer-Phe-CMK, 1 vial, #6149 • Hoechst Stain, 1 mL, #639 • Propidium Iodide, 1 mL, #638 • 10X Cellular Wash Buffer, 15 mL, #6164 • Fixative, 6 mL, #636 • Kit Manual |
Description | FAM-Spacer-Phe-CMK Green FLISP™ Assay Kit |
Supplier | ImmunoChemistry Technologies |
Notes | 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-Spacer-Phe-CMK (FSFCK), 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. |
Product Type | Assay & Detection |
Protocol | 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. FAM-FLISP excites at 488 nm and emits at 520 nm. |
Shipping Information | BLUE ICE |
Size | 25 Tests |
Storage | -20°C, 4°C |
Technical Specifications | • Reagent name: FSFCK • Target: Chymotrypsin-like enzymes • Excitation/Emission: 488 nm / 520 nm • Method of Analysis: Flow cytometry, Fluorescence microscope, fluorescence plate reader • Types of Samples: Cell culture, tissue • Storage: FLISP at ≤ -20°C, all other kit components at 2-8°C • Shipping: Ships overnight (domestic), International Priority Shipping |
Product Page Updated | 2024-01-03T12:33:39.983Z |