Apoptosis Poly Caspases Detection Kits, FLICA and FLIVO

Poly-Caspases Detection Kits
FLICA™: in vitro use with whole cells
FLIVO™: in vivo use in live animals

FLICA™: Quantitate caspase activity (and apoptosis) in whole living cells in vitro

FAM-FLICA Green Poly Caspases Apoptosis Detection Kit # 91
FAM manual
Green FLICA™ kits contain: FAM-FLICA™ reagent, apoptosis wash buffer, fixative, Hoechst 33342, and propidium iodide. Cat.# 91, 25 tests, above.

Poly Caspases FAM-FLICA™ kits
# tests	cat#	price
25	91	$169
100	92	$473

SR-FLICA Red Poly Caspases Apoptosis Detection Kit # 917
SR manual
Red FLICA™ kits contain: SR-FLICA™reagent, apoptosis wash buffer, fixative, and Hoechst 33342. Cat.# 917, 100 tests, above.

Poly Caspases SR-FLICA™ kits
# tests	cat#	price
25	916	$179
100	917	$489

Detect specific caspases with ICT's other green FAM-FLICA™ kits:
Caspase 1
Caspase 2
Caspases 3&7
Caspase 6
Caspase 8
Caspase 9
Caspase 10
Caspase 13

Detect specific caspases with ICT's other red SR-FLICA^™ kits:
Caspases 3&7
Caspase 9

Order today! 1-800-829-3194.

     Use ICT's green or red Poly Caspases FLICA™ kit to quantitate apoptosis via active caspases in whole, living cells. FLICA™ is not an ELISA and does not use any antibodies for detection. Instead, it uses an inhibitor sequence of caspases (VAD) linked to a green (carboxyfluorescein, FAM) or red (sulforhodamine, SR) fluorescent probe. (FLICA™= Fluorescent-Labeled Inhibitor of CAspases.)
     FLICA™ is cell-permeant so you don't have to lyse the cells or permeabilize the membranes. Once added to the culture media, FLICA™ passes through the cell wall. If there is an active caspase enzyme inside the cell, it will form a covalent bond with FLICA™ and retain the fluorescent signal within the cell. Because the caspase enzyme itself binds with FLICA™, there is no interference from pro-caspases nor inactive forms of the enzyme. FLICA™ makes it very easy to identify apoptotic cells.
     Just add FLICA™ to your cell culture media, let it incubate 1-4 hours, wash the cells, and then read. Add a necrotic stain, like PI (which is included in the green FAM kits) or 7-AAD to distinguish apoptotic cells from necrotic cells. To label DNA, Hoechst 33342 is included. Once labeled with FLICA™, cells can be fixed, embedded, or frozen for storage (a fixative is included). Read the fluorescent signal with a microscope, plate reader, or flow cytometer. The green FAM-FLICA™ excites at around 490nm and emits at 530nm (FAM manual); the red SR FLICA™ excites at 560 and emits at 590 (SR manual).
     FLICA™ works with suspension cells, adherent cells, thin tissue sections, and thin frozen sections (list of cell types). FLICA™ will not work with fixed nor paraffin embedded cells, as the caspase enzymes have become inactivated. However, cells can be fixed, embedded, or frozen after labeled with FLICA™. We recommend reading within 24 hours, as the fluorescent label may photobleach, however samples have been frozen for 8 weeks and re-analyzed with equivalent results.
     Poly Caspases FLICA™ kits come in 2 colors: green or red fluorescence; and in 2 sizes: 25 tests and 100 tests. For example, 1 test is a 300uL aliquot of cells grown at 1X10⁶ cells/mL and analyzed on a fluorescence plate reader. Plate readers tend to require the most reagent, flow cytometers the least. Read more about FLICA™, see data, and read protocols:

Don't set up another apoptosis experiment until you call us Order today! 1-800-829-3194.

4 out of 5 cells are apoptotic: Jurkat cells were labeled with ICT's Poly-Caspases FLICA™ kit (cat.# 92). 4 cells fluoresce green (left), but the grey image (right) reveals 5 cells in the field. The 4 green cells are apoptotic = 80% of cells in this experiment had active caspases. The level of fluorescence can be quantified on a fluorescence plate reader or flow cytometer. Data courtesy of Dr. Brian W. Lee, ICT.

Data:

Apoptotic Jurkat cells labeled with Hoechst and Poly Caspases FLICA, catalog # 91.

Jurkat cells dually stained with Hoechst and poly-caspases FLICA™ (catalog #91). Caspase activity is revealed by green fluorescence in the middle cell, indicating that only this cell is apoptotic. Cell at left is also dying (scattered blue), but is not apoptotic because it is not green. Cell in upper right is healthy (concentrated blue nucleus, no green).

List of cell lines.

Apoptosis slide show.

FLICA™ detects neurotoxicity in primary neurons (microscope).

Caspases revealed in yeast using SR-FLICA™ (FACS).

Detect caspases and serine proteases concurrently using flow cytometry.

Stathmoapoptosis in HL60 cells.

Data and protocol in our green FAM-FLICA manual.

Data and protocol in our red SR-FLICAmanual.

Data in our newsletter.

References

FLIVO™: Detect caspase-positive tissues in live animals in vivo

Use FLIVO™ to assess the efficacy of chemotherapy or neurodegeneration in whole animals.

Available in green and red fluorescence.

FAM-FLIVO™ kits contain: FAM-FLIVO™ reagent, and injection buffer.

In Vivo Green FLIVO™ kits
# mice	cat#	price
6	980	$184
24	981	$504

SR-FLIVO™ kits contain: SR-FLIVO™ reagent, and injection buffer.

In Vivo Red FLIVO™ kits

# mice cat# price

6 982 $184

24 983 $504

References

Order today! 1-800-829-3194.

     FLIVO™ reagents are based cell-permeant inhibitors specifically targeted by all active caspases: they are poly-caspase probes that have been optimized for injection into live animals.
     After injection IV, the fluorescent probe (red or green) enters the cell and forms a covalent bond with active caspases; any unbound reagent is washed out of the cell by the natural circulation of the animal. Cells with active caspases fluoresce, non-apoptotic cells do not. No additional processing steps are necessary after sacrifice; simply prepare tissue samples as usual.
     FLIVO™ has successfully labeled tumors, splenocytes, bone marrow, brain, and eye tissues. Use it to assess the efficacy of chemotherapy (did the treatment induce apoptosis?) or monitor neurodegeneration (is the brain dying?). Available in green and red fluorescence. FLIVO™ makes it easy to label apoptosis within the animal. Read a lot more about FLIVO™and see data:

Apoptosis was quantified in excised breast cancer tumors, FLIVO cat.#981 Inject FLIVO into a mouse tail vein IV!

Contol breast cancer tumor has low level of apoptosis, FLIVO cat.#981
apoptosis before treatment

Treated breast cancer tumor has a high level of apoptosis, FLIVO cat.#981
apoptosis after treatment

This drug is working to treat fibrosarcoma tumors.
The level of apoptotic cells significantly increased after treatment (there is an increased level of green fluorescence, below). Data courtesy of Dr. R. Griffin, U of MN, U of AR.

Read the details of this experiment (pdf)

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This page was updated on 01/05/2010 . Please contact Sally Hed with questions 952-888-8788 x10 sally@immunochemistry.com .

APOPTOSIS CASPASE DETECTION KITS FOR IN VITRO (FLICA) AND IN VIVO USE (FLIVO)