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Apoptosis Detection Kits
 Detect apoptosis in live animals in vivo.
Detect caspase activity in whole living cells in vitro.
Monitor apoptosis over time.

 
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More reliable than annexin.
FLICA™ kits are more accurate and reliable than annexin. Annexin detects phosphatidyl serine which is exposed upon the turnover of the cell membrane. This is not an accurate indicator of apoptosis as this turnover can occur for other reasons. Annexin tends to bind to all thymus-derived cells, apoptotic or not. 		Earlier and easier than TUNEL.
 ICT's kits are easier and more sensitive than TUNEL. TUNEL is a long procedure based on DNA laddering. FLICA™ kits detect apoptosis based on caspase activity, which occurs before DNA laddering, so you get an earlier indicator of apoptosis.

In Vivo Apoptosis Detection with FLIVO™
    Researchers can now quantify apoptosis in living animals using ICT’s new FLIVO™ fluorescent in vivo apoptosis detection kits. FLIVO™ is an injectable fluorescent probe used to quantitate apoptosis in live animals. It is a direct stain; once labeled, tissues are ready for analysis and no further processing is necessary. FLIVO™ is very easy to use. Just inject it into the animal and let it circulate 30-45 minutes. Apoptotic cells fluoresce green or red. View cells and tissues directly through a window chamber system        
              control tumor                             treated tumor
or other accessible cavity, or sacrifice the animal and analyze cells with a fluorescent microscope, plate reader, or flow cytometer (see splenocyte data and tissue data). The tissues can be fixed or frozen for future analysis; protect from light during storage and handling.  

 

These pictures were taken of live tumors growing inside 2 mice after injection with red SR-FLIVO™ (kit cat.# 982). The control mouse (left) received a placebo while the test mouse (right) was treated with ATO. 24 hours after treatment, SR-FLIVO™ was given to both mice IV; pictures were taken 30 minutes later. The control tumor exhibits some level of apoptosis as expected, whereas the ATO-treated mouse tumor exhibits a much higher level of apoptosis (it is brighter red).

In Vitro Caspase Detection with FLICA™   

ICT's FLICA™ kits make it easy to measure active caspases in whole living cells: apoptotic cells fluoresce red or green.  Use FLICA™ to quantitate apoptosis, to distinguish apoptosis from necrosis.  

• Easy Just add the reagent directly to the cell culture and incubate.


          normal fibroblasts                        keratoconus
• Fast The reactions start within 15 minutes of addition to the cells, however we recommend an incubation of 1-4 hours. 
• Accurate There is no interference from pro-caspases or inactive forms of the enzyme.
• Reliable Only cells with active caspase enzymes fluoresce. 

Normal (left) and keratoconus (right) corneal fibroblasts were treated with 200uM H2O2 and labeled with FAM-FLICA™ Caspases 3&7 (kit cat.# 93). Treated keratoconus corneal fibroblasts (right) show a significant increase in caspases 3&7 activity compared to treated normal cells (left). Non-apoptotic cells are dark in background.  Data courtesy of Dr. Cristina Kenney, M.D., Ph.D. Dept. of Ophthalmology, UC Irvine

Monitor Apoptosis Over Time with Magic Red™   

 

 

 

 

  To watch apoptosis develop in real time, use ICT's Magic Red kit (#935, $139). This reagent uses a substrate peptide sequence of caspases 3&7, so it does not fluoresce until it’s actually cleaved by active caspases 3&7. We typically watch color develop over several hours, and have presented data over 16 hours. 

 

 

 

 

 

Rat fibroblasts were seeded in 12 well plates at 10,000 cells in 1mL and irradiated the following day. 500uL was removed; 23.1uL of MR-DEVD solution (cat. #935) was mixed with 200uL media and added. 1 hour later, 300uL media was added and cells were photographed over 16 hours. Red fluorescence became brighter as caspase activity and apoptosis progressed. Time 0 is at left, 16 hours is at right.  Data courtesy of Dr. Martin Purschke, MA General Hospital.  View movie clip of cells.

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800-829-3194                                    This page was updated on 04/04/2008 . Please contact us at 952-888-8788.