NIR-FLIVO 690 Free Dye Control Assay
ICT’s near-infrared (NIR)-FLIVO® Tracers are used in conjunction with NIR-FLIVO Free Dye Control Assays. The NIR-FLIVO Free Dye Control Assay uses the NIR-FLIVO Free Dye Control reagent (*Dylight® 690 Free Dye). When injected, both the Free Dye Control and the Tracer reagents will generate a fluorescent signal, but only the Tracer should bind to active caspases and remain inside an apoptotic cell, while signal associated with the Free Dye Control represents background noise. FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
NIR-FLIVO 690 Free Dye Control Assay
FLIVO® kits provide a simple and accurate method to detect caspase activity in vivo. The NIR-FLIVO Free Dye Control Assay contains the NIR fluorescent dye molecule (without the caspase-targeting moiety) and are designed to be used in coordination with the NIR-FLIVO 690 Tracer Assay to determine background fluorescence signal. Both the Free Dye Control and the Tracer reagents will generate a fluorescent signal when injected into animals, but only the Tracer should bind to active caspases and remain inside an apoptotic cell. The fluorescent signal generated from the Free Dye Control reveals where the fluorescent reagent may have become trapped inside the cell, tissues, or body of the animal without specifically binding with an active caspase. In the context of the overall experiment, this base level of fluorescence is considered background noise compared with the signal generated in animals that were injected with the NIR-FLIVO Tracer.
NIR-FLIVO® 690 Free Dye Control reagent (*Dylight® 690 Free Dye)
Background fluorescence levels
690 nm/709 nm
Use instrumentation designed for non-invasive live animal imaging, alternatively, excised tissues can be analyzed by fluorescence microscope or flow cytometer
Whole animal fluorescence imaging, excised tissues
-20°C
Ships overnight (domestic), International Priority Shipping
United States
- Prepare samples and controls.
- Dilute 10X Injection Buffer 1:10 with 45 mL diH20.
- Reconstitute NIR-FLIVO 690 Free Dye with 50 μL DMSO.
- Dilute NIR-FLIVO 690 Free Dye 1:10 with 450 μL 1X Injec¬tion Buffer.
- Inject 50 μL intravenously.
- Let NIR-FLIVO 690 Free Dye circulate for 4 hours.
- Image the animal. NIR-FLIVO 690 Free Dye excites at 690 nm and emits at 709 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.
Kit 9113:
2 vials of NIR-FLIVO® 690 Free Dye Control (*Dylight® 690 Free Dye), 10 Tests per vial, 47.7 μg per vial, #6307
1 bottle of 10X Injection Buffer, 5 mL, #6220
Kit Manual
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
Product Specific References
| PMID | Publication |
| 33627663 | Kolb, R., et al. 2021. Proteolysis-targeting chimera against BCL-XL destroys tumor-infiltrating regulatory T cells. Nature communications, 1281. |