Immunochemistry Technologies, LLC

Quantitate caspase 9 activity inside whole living cells without using antibodies. See See Primary neuron data prices & sizes read the full green procedure in the manual read the full red procedure in the manual see data in our catalog watch our apoptosis slide show Caspases revealed in yeast! data
ICT’s caspase 9 FLICA™ kits measure active caspase 9 inside whole living cells. They are NOT ELISA kits and do NOT use antibodies. ICT has 2 kits to detect caspase 9 - both are FLICA kits - one uses a green fluorescent inhibitor, FAM-LEHD-FMK and another uses a red fluorescent inhibitor, SR-LEHD-FMK. The green FAM FLICA excites at around 490nm and emits at 530nm; What's in the kits? The FAM FLICA™kit contains: the green caspase 9 FLICA™ probe (FAM-LEHD-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). The SR FLICA™kit contains: the red caspase 9 FLICA™ probe (SR-LEHD-FMK); 10x apoptosis wash buffer (which does not lyse the cells); Hoechst stain (a blue DNA stain); and a fixative (which is used after labeling the cells). How do they work? The FLICA™ reagent (Fluorescent Labeled Inhibitors of Caspases) is comprised of 3 segments - it includes: a green (FAM =carboxyfluorescein) or red (SR=sulforhodamine) fluorescent label; an amino acid peptide inhibitor sequence targeted by active caspase 9 (LEHD); and a fluoromethylketone group (FMK) which acts as a leaving group and forms a covalent bond with the active enzyme. The caspase 9 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 9 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; the red SR FLICA™ probe excites at 560 and emits at 590. How many cells do I need? You can detect caspase 9 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; 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 9 enzyme. See our accessories page for a caspase 9 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, 8, 10, or 13) and additional red SR-FLICA™ kits which react with all caspases, and 3&7. Use the red and green kits together and with our serine protease and cholinesterase kits for dual-labeling studies.

FLICA™ Caspase 9 Detection Kits (green & red fluorescent inhibitors)
		25 tests/kit		100 tests/kit
		Cat #	Price	Cat #	Price
9	Green FAM-LEHD-FMK Caspase 9 FLICA™ Kit	912	$169	913	$473
9	Red SR-LEHD-FMK Caspase 9 FLICA™ Kit	960	$179	961	$489

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

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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. 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.
5. inner-Staines, B., VandenHoek, S. 2004. Using cryopreserved neuronal cells for multiwell neurotoxicity assays. Poster presented at the Society for Neuroscience October 2004 annual meeting.
6. Trummell, H.Q., Raisch, K.P., Seay, L.L., and Bonner, James, A. 2004. AKT activity following exposure to IMC-C225 alone, radiation alone or a combination of both treatments. Poster presented at the American Association for Cancer Research, March 27-31, 2004, abstract # 1293.
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 10:31-39.1.
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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