Molecular Formula: N/A

CAS Number: [N/A]

Unit Price (USD): 1 kit $518.70

Bulk Price/unit (when you buy 5 or more): $518.70

Description:
SR-FLICA™ Apoptosis Detection Kits use a novel approach to detect active caspases.

Note:

Application:
The methodology is based on a unique cell-permeable and non-cytotoxic reagent called the Fluorochrome Inhibitor of Caspases (FLICA™). Once inside the cell, the FLICA™ inhibitor binds covalently to the active caspase. The SR FLICA™ reagent excites at 550 nm and has a maximum emission range of 590-600 nm. For the SR FLICA™ kits, which fluoresce red, a sulforhodamine-labeled fluoromethyl ketone peptide inhibitor of caspases is used. The red SR-FLICA™ kits contain a fluorescent-labeled inhibitor, either SR-VAD-FMK (sulforhodaminyl-L-valylalanylaspartyl fluoromethyl ketone), or SR-DEVD-FMK (sulforhodaminyl-L-aspartylglutamylvalyl-aspartylfluoro-methyl ketone). VAD is an amino acid sequence targeted by all caspases, while DEVD is targeted by caspases-3 and -7. When added to a population of cells, the FLICA™ probe enters each cell and covalently binds to a reactive cysteine residue that resides on the large subunit of the caspase heterodimer, thereby inhibiting further enzymatic activity. Because the FLICA™ reagent is covalently coupled to the enzyme, it is retained in the cell, while any unbound FLICA™ reagent will diffuse out of the cell and is washed away. The remaining red fluorescent signal is a direct measure of the number of active caspase enzymes that were present in the cell when the reagent was added. Cells that contain the bound FLICA™ can be analyzed by 96-well-plate based fluorometry, and fluorescence microscopy. Because the FLICA™ reagent SR-VAD-FMK irreversibly binds to many activated caspases (caspases-1, -3, -4, -5, -6, -7, -8 and -9), it can be used as a generic probe for the detection of most caspases. In comparison, when the FLICA™ reagent SR-DEVD-FMK enters the cell, it primarily binds to caspase-3, therefore it can be used to measure the amount of active caspase-3 that was present in the cell at the time when the SR-DEVD-FMK FLICA™ reagent was added. Cells labeled with the FLICA™ reagent may be read immediately or preserved for 24 hours using the fixative. Unfixed SR samples may be analyzed with Hoechst stain.

Product Information Sheet Available Here

References:

• Arends, M.J., Wylie, A.H., (1991) "Apoptosis: Mechanisms and roles in pathology." Int. Rev. Exp. Pathol. 32:223-254.
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• Salvesen, G.S., Dixit, V.M., (1997) "Caspases: intracellular signaling by proteoplysis", Cell 91:443-446.
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• Kumar, S. 1999. Mechanisms mediating caspase activation in cell death. Cell Death and Differ. 6: 1060- 1066.
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• Ekert, P. G., J. Silke and D. L. Vaux. 1999. Caspase inhibitors. Cell Death and Differ. 6: 1081- 1086.