FAM-FLIVO In vivo Poly Caspase Assay
FLIVO® (FLuorescence in vIVO) is a powerful method for assessing caspase activity in vivo. FAM-FLIVO poly caspase probes are non-cytotoxic, cell-permeant fluorescent inhibitors of caspases optimized for use in whole live animals. ICT’s FAM-FLIVO® poly caspase inhibitor probe contains the preferred binding sequence for most caspases (Val-Ala-Asp or VAD). This preferred poly caspase tripeptide binding sequence is labeled with a carboxyfluorescein (FAM) dye and a fluoromethyl ketone (FMK) reactive entity.
FAM-FLIVO In vivo Poly Caspase Assay
FLIVO® kits provide a simple and accurate method to detect caspase activity in vivo. Similar to our FLICA® probes but optimized for whole live animal labeling. To label cells containing active caspases, inject FLIVO intravenously and let it circulate ~60 minutes. Because the reagent is cell-permeant, it readily diffuses in and out of all cells that it encounters as it circulates throughout the body. If there are active caspase enzymes inside a cell, FLIVO will form an irreversible covalent bond with the caspase active site. The bound FLIVO probe will remain inside the cell if the cell membrane is intact. Any unbound FLIVO is removed from the circulation of the animal in about an hour. The remaining green fluorescent signal in the tissue is a direct measure of caspase activity that occurred at the time the reagent was injected. Once the excess FLIVO has cleared from the body of the animal, the tissues are ready for analysis. No further staining is necessary. Tissues can be viewed directly through a window chamber system or other accessible cavity. Alternatively, target tissues can be removed and processed for analysis. FLIVO is very sensitive and will pick up naturally occurring background apoptosis. Apoptotic cells have more active caspases than control cells, therefore they fluoresce brighter with FLIVO.
FAM-FLIVO® Poly Caspase Inhibitor (FAM-VAD-FMK)
Poly Caspases
492 nm/520 nm
Fluorescence Microscope, Flow Cytometer, Window Chamber System
Whole live animal, excised tissue
2-8°C
Ships overnight (domestic), International Priority Shipping
United States
- Prepare samples and controls.
- Dilute 10X Injection Buffer 1:10 by adding 45 mL diH20.
- Reconstitute FAM-FLIVO with 50 µL DMSO.
- Dilute FAM-FLIVO 1:12 with 550 µL 1X Injection Buffer.
- Inject 100 µL intravenously.
- Let FAM-FLIVO circulate 30-60 minutes.
- View live tumor through a window chamber using a fluorescence microscope.
- If not viewing directly, excise tissue.
- If desired, label with additional stains, such as Hoechst 33342, or an antibody.
- If desired, fix cells.
- Analyze with a fluorescence microscope, flow cytometer, or a window chamber system. FAM-FLIVO excites at 492 nm and emits at 520 nm.
Kit 980 6 Tests:
FAM-FLIVO® Poly Caspase Inhibitor (FAM-VAD-FMK), 1 vial, #6218
10X Injection Buffer, 5 mL, #6220
Kit Manual
Kit 981 24 Tests:
FAM-FLIVO® Poly Caspase Inhibitor (FAM-VAD-FMK), 4 vials, #6218
10X Injection Buffer, 5 mL, #6220
Kit Manual