Red Sulforhodamine Poly Caspases Detection Kit 100tests

Product ID: M0834



Unit SizePriceQuantity 
1kit
$599.71

Availability: In stock


Description

SR-FLICA® Apoptosis Detection Kits use a novel approach to detect active caspases.

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 SR-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 SR-FLICA® reagent is covalently coupled to the enzyme, it is retained in the cell, while any unbound SR-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 SR-FLICA® can be analyzed by 96-well-plate based fluorometry, and fluorescence microscopy. Because the SR-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 SR-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 SR-FLICA® reagent was added. Cells labeled with the SR-FLICA® reagent may be read immediately or preserved for 24 hours using the fixative. Unfixed SR samples may be analyzed with Hoechst stain.

Technical Data
SKU M0834
Unit Size 1kit
Detection Method Fluorescence

References and Citations

References:

  • Arends, M.J., Wylie, A.H., (1991) "Apoptosis: Mechanisms and roles in pathology." Int. Rev. Exp. Pathol. 32:223-254.
  • Slee, E.A., Adrian, C., Martin, S.J., (1999) "Serial Kollers: Ordering Caspase Activation events in Apoptosis", Cell Death and Differ. 6: 1067-1074
  • Salvesen, G.S., Dixit, V.M., (1997) "Caspases: intracellular signaling by proteoplysis", Cell 91:443-446.
  • 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 specificities 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.
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