Magic Red™ Caspases 3 & 7 Detection Kit

Assess caspases 3&7 activity in vitro and in real time

Magic Red-DEVD kits measure apoptosis via active caspases 3&7 in whole living, intact cells - no lysis required. As apoptosis progresses and caspase activity increases, the red fluorescent signal increases. These unique kits are not ELISAs and do not use antibodies - instead they use a cell-permeant substrate sequence, DEVD, specifically targeted by active caspases 3 & 7. This peptide sequence is linked to a red fluorophore, Magic Red, which fluoresces once cleaved by active caspases 3 or 7.  This kit is a good assay to monitor apoptosis over time. 

Just add the reagent to media, let it incubate, and read the cells.  If there are active caspase 3&7 enzymes, the reagent will fluoresce red, if not, it will not fluoresce.  Watch 2 movies of real-time caspase activity, courtesy of Dr. M. Purschke, MA General Hospital: movie clip of cells ; movie of fluorescence. ICT’s MR-DEVD kits give a clear picture of apoptotic versus non-apoptotic cells and enable you to monitor caspase activity over time.

Caspases 3&7 are active in the cytosol. After staining with MR, there are often areas of bright red fluorescence located within lysosomal structures. However, caspases are not known to be present within lysosomes.  The presence of the red fluorescent product within the lysosomes is most likely due to the migration and concentration of the cleaved red fluorescent fluorophore from the cytosol into the lysosomes, thereby creating bright spots.

 

Magic Red Caspases 3&7 Detection kit contains:

  • ICT's red Magic Red caspases 3&7 detection probe, MR-(DEVD)2
  • Hoechst stain (a blue DNA stain)
  • Acridine orange (a lysosomal stain) 
  • MR-Caspases 3&7 Apoptosis Manual

Trial size, ~25 tests:

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Regular size, ~100 tests:

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Learn more at ICT's version 1.0 Magic Red webpage

Inhibitor-based kits (FLICA)

In addition to Magic Red, ICT also offers inhibitor-based kits to detect Caspases 3&7 in whole cells. These kits are available in red or green fluorescence.

Magic Red image

ICT's MR Caspases 3&7 kit clearly distinguishes apoptotic MCF-7 cells from non-apoptotic cells. Apoptotic cells exhibit red lysosomes (MR) with less intense blue nuclei. These cells are intermixed with non-apoptotic cells bearing bright blue nuclei (Hoechst) and absent lysosomal staining.

Sample protocol:

  1. Culture your cells up to 1 x 10^6 cells/mL.
  2. Induce apoptosis following your protocol, and create positive and negative controls.
  3. Reconstitute the reagent with DMSO to form the stock concentrate (which can be frozen for future use).
  4. Dilute the stock concentrate with 1X PBS to form the working solution.
  5. Add ~10mcL of the working solution directly to a 300-500mcL aliquot of your cell culture for labeling.
  6. Incubate 1-4 hours.
  7. If desired, label DNA with Hoechst stain.
  8. If desired, label lysosomes with Acridine orange.
  9. If desired, fix cells.
  10. Analyze data using a fluorescence microscope, or plate reader.