Immunochemistry Technologies, LLC

Quantitate caspase 8 activity inside whole living cells without using antibodies.

prices & sizes
FAM-FLICA manual
data in our catalog
apoptosis slide show
Caspases revealed in yeast! data

     ICT’s caspase 8 FLICA™ kit measures active caspase 8 inside whole living cells. It is NOT an ELISA and does NOT use antibodies.

What's in the kit?
     The kit contains: the caspase 8 FLICA™ probe (FAM-LETD-FMK); 10x apoptosis wash buffer (which does not lyse the cells); Hoechst stain (a blue DNA stain); propidium iodide (a red live/dead stain); and a fixative (which is used after labeling the cells).

How does it work?
     The FLICA™ reagent (Fluorescent Labeled Inhibitors of Caspases) is comprised of 3 segments - it includes: a green (FAM =carboxyfluorescein) fluorescent label; an amino acid peptide inhibitor sequence targeted by active caspase 8 (LETD); and a fluoromethylketone group (FMK) which acts as a leaving group and forms a covalent bond with the active enzyme.
     The caspase 8 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 8 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 .

How do I read my cells?
     The resulting positive fluorescent signal can be detected with a fluorescence

microscope (see catalog for data), a fluorescence plate reader, or a flow cytometer. The green FLICA™ probe, FAM, excites at 490nm and emits at 520nm.

How many cells do I need?
     You can detect caspase 8 activity in individual cells or culture them up to 1x10⁶ cells/mL and read them using a fluorescence plate reader or flow cytometer. They also work with adherent cells.

What kind of cells can I use?
     FLICA™ kits have been used successfully with many types of cancer cells; primary cells, neurons, glial 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 8 enzyme. See our accessories page for a caspase 8 antibody and other reagents.

What other kits do you have?
     To detect apoptosis in general, use the green or red poly caspases kits. These kits use a general peptide sequence (VAD) which reacts with all active caspases. To target other specific caspase, ICT offers several green FAM-FLICA™ kits which react with different caspases (caspase 1, 2, 3&7, 6, 9, 10, or 13) and red SR-FLICA™ kits which react with all caspases, 3&7, and caspase 9. Use the red and green kits together and with our serine protease and cholinesterase kits for dual-labeling studies.

Order today! Just call 1-800-829-3194

or download our simple order sheet (Word) or PDF
main: 952-888-8788
fax: 952-888-8988

FLICA™ Caspase 8 Detection Kit (green fluorescent inhibitor, FAM-LETD-FMK)
Caspase	Green FLICA™ Kits	25 tests/kit		100 tests/kit
		Cat #	Price	Cat #	Price
8	FAM-LETD-FMK Caspase 8 FLICA™ Kit	99	$169	910	$473

To get your free t-shirt, just order any 2 products from this website or our newsletter and mention code VOL6 when placing the order. It's that easy! (Sorry, offer does not apply to orders placed with most distributors.)

If you have any questions about your order, please contact Sally Hed at 800-829-3194 or 952-888-8788 or sally@immunochemistry.com .

References
1. Grabarek, J., P. Amstad, and Z. Darzynkiewicz. 2002. Use of fluorescently labeled caspase inhibitors as affinity labels to detect activated caspases. Human Cell vol 15(1):1-12.
2. Grabarek, J., and Z. Darzynkiewicz. 2002. In situ activation of caspases and serine proteases during apoptosis detected by affinity labeling their enzyme active centers with fluorochrome-tagged inhibitors. Exp. Hematology vol 30:982-989.
3. Grunewald, S., Paasch, U., Said, T.M., Sharma, R.K., Glander, H.J., Agarwal, A. 2005. Caspase activity in human spermatozoa in response to physiological and pathological stimuli. Fertility and Sterility vol 83:1106-1112.
4. Kemp, T.J., Ludwig, A.T., Earel, J.K., Moore, J.M., VanOosten, R.L., Moses, B., Leidal, K., Nauseef, W.M. and Griffith, T.S. 2005. Neutrophil stimulation with Mycobacterium bovis bacillus Calmette-Guerin (BCG) results in the release of functional soluble TRAIL/Apo-2L. Blood vol 106(10):3474-3482
5. Paasch, U., Sharma, R.K., Gupta, A.K., Grunewald, S., Mascha, E.J., Thomas, A.J. Jr., Glander, H.J., and Agarwal, A. 2004. Cryoperservation and thawing is associated with varying extent of activation of apoptotic machinery in subsets of ejaculated human spermatozoa. Biology of Reproduction vol 71:1828-1837.
6. Tatari-Calderone, Z., Brogdon, J.L., Tinsley, K.W., Ramezani, A., and Leitenberg, D. 2005. CD4-dependent signaling is required for a late checkpoint during Th2 development associated with resistance to activation-induced cell death. The Journal of Immunology vol 175:5629-5636
7. VanOsten, R.L., Moore, J.M., Karacay, B. Griffith, T.S. 2005. Histone deacetylase inhibitors modulate renal cell carcinoma sensitivity to TRAIL/Apo-2L-induced apoptosis by enhancing TRAIL-R2 expression. Cancer Biology & Therapy vol 4, issue 10:31-39.
8. Wang, Z., Goulet, R. 3rd, Stanton, K.J., Sadaria, M., and Nakshatri, H. 2005. Differential effect of anti-apoptotic genes Bcl-xL and c-FLIP on sensitivity of MCF-7 breast cancer cells to paclitaxel and docetaxel. Anticancer Res vol 25(3c):2367-2379.

See all FLICA references (pdf)

Order today! Just call 1-800-829-3194

or download our simple order sheet (Word) or PDF
main: 952-888-8788
fax: 952-888-8988

Please contact Sally Hed with questions 952-888-8788 x10 sally@immunochemistry.com
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