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.
- 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.
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