FLIVO® (FLuorescence in vIVO) is a powerful method for non-invasive detection of caspase activity in vivo. NIR-FLIVO 747 Tracers are non-cytotoxic, cell-permeant fluorescent inhibitors of caspases optimized for use in whole live animals. ICT’s NIR-FLIVO 747 Tracer probe contains the preferred binding sequence for most caspases (Val-Ala-Asp or VAD). This preferred poly caspase tripeptide binding sequence is labeled with *Dylight® 747 dye and a fluoromethyl ketone (FMK) reactive entity. FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
- Prepare samples and controls.
- Dilute 10X Injection Buffer 1:10 with 45 mL diH20.
- Reconstitute NIR-FLIVO 747 Tracer with 50 μL DMSO.
- Dilute NIR-FLIVO 1:10 with 450 μL 1X Injection Buffer.
- Inject 50 μL intravenously.
- Let NIR-FLIVO circulate for 4 hours.
- Image the animal. NIR-FLIVO 747 Tracer excites at 747 nm and emits at 776 nm.
- If not viewing directly, excise tissue and process samples for further analysis.
- Optional: label excised samples with additional stains, such as Hoechst 33342, or an antibody.
- If desired, fix excised samples.
- Analyze excised samples with a fluorescence microscope or flow cytometer.
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
Wu Q, Berglund AE, Wang D, MacAulay RJ, Mulé JJ, Etame AB. Paradoxical epigenetic regulation of XAF1 mediates plasticity towards adaptive resistance evolution in MGMT-methylated glioblastoma. Sci Rep. 2019 Oct 1;9(1):14072. doi: 10.1038/s41598-019-50489-2. Full Text