FLICA™ Caspase 10 Detection Kit

Quantitate caspase 10 in vitro with FAM-AEVD-FMK

ICT's FLICA caspase 10 detection assay is a simple yet accurate method to measure caspase 10 activity in whole cells. Simply add the FLICA caspase 10 reagent to the media and incubate. Cells containing active caspase 10 will fluoresce green. The kit contains:

Trial size, ~25 tests:
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Regular size, ~100 tests:
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Caspase 10 activity may be detected in individual cells or suspensions of up to 1x10^6 cells/mL. The noncytotoxic FLICA probe is cell permeant: no lysis or permeabilization steps are necessary. Just culture cells, add FLICA to the media, incubate, and wash. Active caspase 10 enzymes will bind to FLICA and retain it within the cells, thereby capturing the green fluorescent signal. FLICA constantly fluoresces, therefore any unbound reagent must be washed out of the cell to remove any background noise. The resulting positive fluorescent signal can be detected with a fluorescence microscope, a fluorescence plate reader, or a flow cytometer. The green FLICA probe, FAM, excites at 490nm and emits at 520nm.

The FLICA assay also works with adherent cells and very thin tissue sections. FLICA will not work in previously fixed cells or paraffin cells, as those treatments inactivate the caspase 10 enzyme. Please call for details or technical questions at 800-829-3194.

ICT's caspase 10 inhibitor probe is comprised of 3 segments:

FLICA kits have been used successfully with a multitude of cell lines from a variety of species (human, mouse, rat, etc), including, but not limited to, cancer cells, neurons, primary cells, epithelial cells, endothelial cells, and macrophages. For in vitro apoptosis detection via active poly caspases, see our FLICA Poly Caspases Detection Kits.

The caspase 10 FLICA reagent is cell permeant, so you don’t have to lyse or permeabilize the membrane to get it into the cell. Just culture your cells, add the FLICA probe to the media, incubate, and wash the cells. If there are active caspase 10 enzymes, they will bind to FLICA and retain it within the cell, thereby capturing the fluorescent signal. The FLICA probe constantly fluoresces, therefore any unbound reagent must be washed back out of the cell to remove any background noise.  This is done by several quick rinse and spin steps, or further incubation with the wash buffer or cell culture media. Click here to read our complete manual.

FLICA kits have been used successfully with many types of cancer cells; epithelial cells; endothelial cells; macrophages; and other types of human, mouse, rat tissues, and many other species.

It will not work in previously fixed cells or paraffin cells, as that treatment inactivates the caspase 10 enzyme.  See our accessories page for a caspase 10 antibody and other reagents.

Caspase 10 data 

Jurkat cells were treated with staurosporine, an apoptosis-inducing agent (bottom), or DMSO, a negative control (top), for 4 hours, then labeled with ICT’s green caspase 10 inhibitor probe, FAM-AEVD-FMK, for 1 hour. Cells were washed twice and read on a flow cytometer. Treatment with staurosporine induced caspase 10 activity in 93.6% of the experimental cells (M2, bottom right), whereas the negative control treatment exhibited caspase 10 activity in only 4.7% of the cell population (M2, top right). This is a ratio of 20:1. (Dr. Brian W. Lee, ICT).

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 50mcL DMSO to form the stock concentrate (which can be frozen for future use).
  4. Dilute the stock concentrate with 200mcL 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. Wash and spin cells twice, or let incubate for 1 hour with fresh media or 1x apoptosis wash buffer.
  8. If desired, label cells with Hoechst stain.
  9. If desired, label cells with Propidium Iodide or 7AAD.
  10. If desired, fix cells.
  11. Analyze data using a fluorescence microscope, plate reader, or flow cytometer.