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| Quantitate
caspase 1 activity inside whole living cells without using
antibodies.
prices
& sizes
FAM-FLICA™
manual
data in our newsletter
apoptosis slide show
Caspases revealed in yeast! data
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ICT’s Caspase 1 FLICA™ kit measures active caspase 1
inside whole living cells. It is NOT an
ELISA and does NOT use antibodies.
What's
in the kit?
The kit contains: the
caspase 1 FLICA™ probe (FAM-YVAD-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 1 (YVAD); and a fluoromethylketone group (FMK) which acts as a leaving group and forms a covalent bond with the active enzyme.
The caspase 1 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 1 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 |
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microscope
(see newsletter
Figures 2, 3, 5a, 9), a fluorescence plate reader (Figure
10), or a flow cytometer
(Figure 7). The green FLICA™ probe, FAM, excites at 490nm and emits at 520nm.
How
many cells do I need?
You can detect caspase 1 activity in individual cells (Figures 1, 2, & 6) or culture them up to
1x106 cells/mL and read them using a fluorescence plate reader or flow cytometer (Figures
10 & 7). They also work with adherent cells (Figure
3).
What
kind of cells can I use?
FLICA™ kits have been used successfully with
many types of cancer cells; neurons, primary
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 1 enzyme. See our accessories
page for a caspase 1 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
2,
3&7,
6,
8,
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 (Figures
5, 7, & 9).
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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FLICATM
Caspase 1 Detection Kit
(green fluorescent inhibitor, FAM-YVAD-FMK) |
|
Caspase
|
Green
FLICA™
Kits |
25
tests/kit |
100
tests/kit |
| Cat
# |
Price |
Cat
# |
Price |
| 1 |
FAM-YVAD-FMK
Caspase 1 FLICA™ Kit |
97 |
$164 |
98 |
$459 |
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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
.
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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. Scotton, C.J., Martinez, F.O., Smelt,
M.J., Sironi, M., Locati, M., Mantovani, A., and Sozzani, S. 2005. Transctiptional profiling reveals complex regulation of the monocyte IL-1â system by IL-13. The Journal of Immunology. 834-845.
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
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