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FAM FLICA™ Apoptosis Detection Kits use a novel approach to detect active caspases intracellularly.
The methodology is based on the unique cell-permeable and non-cytotoxic reagents called the Fluorochrome Inhibitor of Caspases (FLICA). Once inside the cell, the FLICA™ inhibitor binds covalently to the active caspase. For the FAM FLICA™ kits, which fluoresce green, a carboxyfluorescein-labeled fluoromethyl ketone peptide inhibitor of the specific caspase is used (EX:488nm / EM:520nm), compatible with most fluorescence microscopes or microplate readers.
STAINING PROTOCOL Staining apoptotic cells with the FLICA™ kit can be completed within a few minutes. However, the FLICA™ kit is used with living cells, which require periodic maintenance and cultivation several days in advance. In addition, once the proper number of cells has been cultivated, time must be allotted for the induction process. The FLICA™ kit works with your current apoptosis protocols - induce apoptosis as you normally would and then label the cells with FLICA™. 1. Culture cells to a density optimal for apoptosis induction according to your specific induction protocol, but not to exceed 106 cells/mL. 2. At the same time, culture a non-induced negative control cell population at the same density as the induced population for every labeling condition. 3. Induce apoptosis following your protocol (such as treating Jurkat cells with 2 mg/ml camptothecin for 3 hours). 4. Dilute the 10X wash buffer to 1X. 5. Reconstitute the vial of lyophilized FLICA™ with DMSO to form the 150X FLICA™ stock concentrate. 6. Dilute the 150X FLICA™ stock to the 30X working solution. 7. Stain cells by adding the 30X FLICA™ solution. 8. Incubate for 1 hour. 9. Wash and spin cells. 10. If desired, label cells with Hoechst 33342 stain. 11. If desired, fix cells. 12. Analyze data via microtiter plate fluorometry or fluorescence microscopy.
Product Information Sheet available here
View References ▼
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- Walker, N. P., R. V. Talanian, K. D. Brady, L. C. Dang, N. J. Bump, C. R. Ferenz, S. Franklin, T. Ghayur, M. C. Hackett and L. D. Hammill. 1994. Crystal Structure of the Cysteine Protease Interleukin-
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- Rotonda, J., D. W. Nicholson, K. M. Fazil, M. Gallant, Y. Gareau, M. Labelle, E. P. Peterson, D. M. Rasper, R. Ruel, J. P. Vaillancourt, N. A. Thornberry and J. W. Becker. 1996. The three- dimensional structure of apopain/ CPP32, a key mediator of apoptosis. Nature Struct. Biol. 3( 7): 619- 625.
- Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060- 1066.
- Thornberry, N. A., T. A. Rano, E. P. Peterson, D. M. Rasper, T. Timkey,M. Garcia- Calvo, V. M. Houtszager, P. A. Nordstrom, S. Roy, J. P. Vaillancourt, K. T. Chapman and D. W. Nicholson. 1997. A combinatorial approach defines pecificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272( 29): 17907- 17911.
- Ekert, P. G., J. Silke and D. L. Vaux. 1999. Caspase inhibitors. Cell Death and Differ. 6: 1081- 1086.
Unit Price (USD):
1 kit $186.33
(5 or more):