NIR-FLIVO® 690 Free Dye Control Assay

$269.00

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.

Catalog # Product Size Quantity Price
9113 20 Tests $269.00
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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.

1. Prepare samples and controls.
2. Dilute 10X Injection Buffer 1:10 with 45 mL diH20.
3. Reconstitute NIR-FLIVO 690 Free Dye with 50 μL DMSO.
4. Dilute NIR-FLIVO 690 Free Dye 1:10 with 450 μL 1X Injec¬tion Buffer.
5. Inject 50 μL intravenously.
6. Let NIR-FLIVO 690 Free Dye circulate for 4 hours.
7. Image the animal. NIR-FLIVO 690 Free Dye excites at 690 nm and emits at 709 nm.
8. If not viewing directly, excise tissue and process samples for further analysis.
9. Optional: label excised samples with additional stains, such as Hoechst 33342, or an antibody.
10. If desired, fix excised samples.
11. Analyze excised samples with a fluorescence microscope or flow cytometer.

DETECTION OF APOPTOSIS IN MURINE BRAIN ABSCESSES USING NIR-FLIVO 690 TRACER
Brain abscesses were induced in mice following the intracerebral inoculation of live Staphylococcus aureus. Animals received intravenous injections of NIR-FLIVO 690 Tracer (kit #9112) or NIR-FLIVO 690 Free Dye Control (kit #9113) at 17 hours post-infection, whereupon signals were acquired 1 hour later from brain tissues ex vivo using an IVIS® Spectrum™ (Caliper Life Sciences). Strong caspase activity was associated with brain abscesses following administration of the NIR-FLIVO 690 Tracer (right image), whereas minimal signal was detected in animals injected with the NIR-FLIVO 690 Free Dye Control (left image).
690 Tracer and Free Dye data

• Reagent Name: NIR-FLIVO® 690 Free Dye Control reagent (*Dylight® 690 Free Dye).
• Target: Background fluorescence levels
• Excitation/Emission: 690 nm/709 nm
• Method of Analysis: Use instrumentation designed for non-invasive live animal imaging, alternatively, excised tissues can be analyzed by fluorescence microscope or flow cytometer.
• Types of Samples: Whole animal fluorescence imaging, excised tissues
• Storage: -20°C
• Shipping: Ships overnight (domestic), International Priority Shipping
FLIVO® is a registered trademark of ICT. *Dylight® is a registered trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

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.