NIR-FLIVO® 747 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® 747 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
9115 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 747 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 747 Free Dye with 50 μL DMSO.
4. Dilute NIR-FLIVO 747 Free Dye 1:10 with 450 μL 1X Injec¬tion Buffer.
5. Inject 50 μL intravenously.
6. Let NIR-FLIVO 747 Free Dye circulate for 4 hours.
7. Image the animal. NIR-FLIVO 747 Free Dye excites at 747 nm and emits at 776 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.

NON-INVASIVE IMAGING OF APOPTOSIS RESULTING FROM HSV-1 VIRAL INFECTION
Adult wild-type Balb/c mice were either inoculated with HSV-1 virus, which is known to induce apoptosis in the brain, or given a sham treatment. Seven days after viral inoculation, the mice were injected intravenously with either the NIR-FLIVO 747 Tracer (kit #9114) to detect caspase activity, the NIR-FLIVO 747 Free Dye (kit #9115) to detect where the dye reagent is getting trapped within tissues, or no reagent. The animals were then imaged with a Carestream In Vivo FX PRO imager seven hours after reagent injection. Strong caspase activity was located in the brain of the animal treated with HSV-1 and injected with NIR-FLIVO 747 Tracer (NIR-FLIVO 747 +). Minimal signal was detected in the liver region of the HSV-treated animal injected with the free dye control (HSV-treated; NIR 747 Free Dye +) and of the uninfected mouse injected with NIR-FLIVO 747 Tracer. The liver is the route of clearance for FLIVO Tracers. All mice show fluorescence signal in the tail where reagent is likely to pool after IV injection.

Apoptosis Imaged in HSV-1+ Mice with NIR-FLIVO 747

• Reagent Name: NIR-FLIVO® 747 Free Dye Control reagent (*Dylight® 747 Free Dye).
• Target: Background fluorescence levels
• Excitation/Emission: 747 nm/776 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 9115:
• 2 vials of NIR-FLIVO® 747 Free Dye Control (*Dylight® 747 Free Dye), 10 Tests per vial, 53.6 μg per vial, #6309
• 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.