Nano-Glo® Fluorofurimazine In Vivo Substrate (FFz)

Reagent Optimized for in vivo Detection

NanoLuc® luciferase provides a new reporter option for advanced in vivo imaging applications. Its small size is ideal for engineering into compact genomes, and ATP-independence allows in vivo monitoring of both intracellular and extracellular events. Substrate specificity makes NanoLuc® luciferase an ideal complementary reporter to firefly luciferase for an improved dual-luciferase in vivo imaging solution.

The Nano-Glo® Fluorofurimazine In Vivo Substrate (FFz) is an optimized reagent designed specifically for in vivo detection of NanoLuc® luciferase, NanoLuc® fusion proteins or reconstituted NanoBiT® luciferase. This optimized, aqueous-soluble, in vivo detection reagent provides increased substrate bioavailability for bright, stable signals and offers flexible delivery options with handling requirements compatible with in vivo workflows.

Ideal for in vivo analysis of:

Dual-reporter imaging with firefly luciferase
Viral distribution
CAR T-cell tracking

Exosome/extracellular vesicles
Optogenetics
BRET-based reporters for enhanced deep-tissue imaging

Flexible Substrate Delivery

The Nano-Glo® Fluorofurimazine In Vivo Substrate can be delivered via multiple routes of administration. Each method offers different signal intensity and kinetics, allowing flexibility between experimental systems.

Mice injected with FFz via i.p. injection

Mice injected with FFz via i.v. injection

Graph comparing FFz i.p. and i.v. injection methods

NanoLuc® signal of 4T1 primary tumors in BALB/c mice. Cells expressing NanoLuc® luciferase were orthotopically implanted into the #3 mammary fat pad of female BALB/c mice, and tumors were allowed to grow for 22 days. Mice were injected with FFz via i.p. (top image; circles) or i.v. injection (bottom image; squares). Bioluminescence imaging was done at the University of Wisconsin Small Animal Imaging and Radiotherapy Facility.

Ideal for Cell and Gene Therapy Applications

The small size of NanoLuc® luciferase makes it easy to package into viral genomes, allowing you to track tissue penetration of your virus for infectious disease or gene therapy applications.

Mice injected with increasing levels of AAV9-NanoLuc®-HaloTag® viral particles

In vivo whole-body imaging of mice injected with AAV9-NanoLuc®-HaloTag® viral particles. Mice were injected with increasing levels of AAV9-NanoLuc®-HaloTag® viral particles. Nano-Glo® FFz (0.44μmol) was injected 7- and 13-days post-transduction followed by in vivo imaging. Tissue tropism starts to appear at 7 days, and a stronger signal and more extensive tissue tropism is visible after 13 days. Bioluminescence imaging was done at the University of Wisconsin Small Animal Imaging and Radiotherapy Facility.

Image the Brain of Living Animals with NanoLuc® Luciferase

Nano-Glo® Cephalofurimazine (CFz9) In Vivo Substrate enables the detection of NanoLuc® Luciferase in the brain of living animals. CFz9 was designed to cross the blood-brain barrier, enabling noninvasive exploration of central nervous system physiology and anatomy and studies of therapeutic distribution to the brain.

Request early access to CFz9 In Vivo Substrate.

Learn more about the CFz9 substrate in this webinar.

Learn More About Imaging with NanoLuc® Luciferase

NanoLuc® reporters can be multiplexed with firefly reporters to enable two-population bioluminescent imaging in animals. This publication used a NanoLuc® reporter to track tumor size and a firefly substrate to visualize CAR-T cells in the same animal over multiple days:

Novel NanoLuc® substrates enable bright two-population bioluminescence imaging in animals

(Su, Y. et al. (2020) Nature Methods 17, 852–860.)

Nano-Glo® Fluorofurimazine In Vivo Substrate (FFz) Citations by Application

Expand the section below to see a list of articles citing use of the Nano-Glo® Fluorofurimazine In Vivo Substrate in a variety of applications.

Expand to View Citations

Here is a collection of articles demonstrating the use of the Nano-Glo® Fluorofurimazine In Vivo Substrate by application. You can search or scroll to find your application.

Search citations

Application	Citation
NanoLuc® Reporter Virus	Ullah, I. et al. (2021) Live imaging of SARS-CoV-2 infection in mice reveals that neutralizing antibodies require Fc function for optimal efficacy. Immunity 54, 2143–58.
NanoLuc® Reporter Virus	Tran, V. et al. (2013) Highly sensitive real-time in vivo imaging of an influenza reporter virus reveals dynamics of replication and spread. J. Virol. 87, 13321–9.
NanoLuc® Reporter Virus	Carossino, M. et al. (2021) Fatal neuroinvasion and SARS-CoV-2 tropism in K18-hACE2 mice is partially independent on hACE2 expression. Viruses 14, 535.
NanoLuc® Reporter Virus	Kim, J.H. et al. (2022) Real-time tracking of bioluminescent influenza A virus infection in mice. Sci. Rep. 12, 3152.
NanoLuc® Reporter Virus	Hollander, J.M. et al. (2023) Adeno-associated virus-delivered fibroblast growth factor 18 gene therapy promotes cartilage anabolism. Cartilage epub ahead of print.
NanoLuc® Reporter Virus	Davies, K.A. et al. (2023) Fluorescent and bioluminescent reporter mouse-adapted Ebola viruses maintain pathogenicity and can be visualized in vivo. J. Infect. Dis. epub ahead of print.
NanoLuc® Reporter Virus	Kanai, Y. et al. (2019) In vivo live imaging of oncolytic mammalian Orthoreovirus expressing NanoLuc luciferase in tumor xenograft mice. J. Virol. 93, e00401-19.
HiBiT Oncolytic Virus	Gasper, N. et al. (2020) NanoBiT System and hydrofurimazine for optimized detection of viral infection in mice—a novel in vivo imaging platform. Int. J. Mol. Sci. 21, 5863–73.
CAR T-Cell Tracking	Theruvath, J. et al. (2020) Locoregionally administered B7-H3-targeted CAR T cells for treatment of atypical teratoid/rhabdoid tumors. Nat. Med. 26, 712–9.
CAR T-Cell Tracking	Su, Y. et al. (2020) Novel NanoLuc substrates enable bright two-population bioluminescence imaging in animals. Nature Methods 17, 852–860.
CAR T-Cell Tracking	Wang, T. et al. (2023) Visualizing cell-cell communication using synthetic notch activated MRI. Proc. Natl. Acad. Sci. 120, e2216901120.
Exosomes/Extracellular Vesicles	Luo, W. et al. (2020) Spatial and temporal tracking of cardiac exosomes in mouse using a nano-luciferase-CD63 fusion protein. Commun. Biol. 3, 114.
Exosomes/Extracellular Vesicles	Gupta, D. et al. (2020) Quantification of extracellular vesicles in vitro and in vivo using sensitive bioluminescent imaging. J. Extracell. Vesicles 9, e1800222.
Exosomes/Extracellular Vesicles	Rufino-Ramos, D. et al. (2022) Using genetically modified extracellular vesicles as a non-invasive strategy to evaluate brain-specific cargo. Biomaterials 281, e121366.
Exosomes/Extracellular Vesicles	Perez, G.I. et al. (2022) In vitro and in vivo analysis of extracellular vesicle-mediated metastasis using a bright, red-shifted bioluminescent reporter protein. Adv. Genet. 3, e2100055.
Cell Tracking	Wang, L. et al. (2021) Application of bioluminescence resonance energy transfer-based cell tracking approach in bone tissue engineering. J. Tissue Eng. 12, e2041731421995465.
Receptor Binding	Tang, Y. et al. (2019) A bioluminescence resonance energy transfer-based approach for determining antibody-receptor occupancy in vivo. iScience 15, 439–51.
NanoLuc® Reporter RNA	Chen, R. et al. (2023) Engineering circular RNA for enhanced protein production. Nat. Biotechnol. 41, 262–72.
NanoLuc® Reporter RNA	Hall, R.N. et al. (2022) Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection. Cell Rep. Methods 2, e100298.
Optogenetics	He, L. et al. (2021) Optogenetic control of non-apoptotic cell death. Adv. Sci. 13, e2100424.
Optogenetics	Li, T. et al. (2021) A synthetic BRET-based optogenetic device for pulsatile transgene expression enabling glucose homeostasis in mice. Nat. Commun. 12, 615.
Deep Tissue BLI	Gaspar, N. et al. (2021) Evaluation of NanoLuc substrates for bioluminescence imaging of transferred cells in mice. J. Photochem. Photobiol. B. 216, e112128.
Deep Tissue BLI	Driedonks, T. et al. (2022) Pharmacokinetics and biodistribution of extracellular vesicles administered intravenously and intranasally to Macaca nemestrina. J. Extracell. Biol. 1, e59.

Protocols

Complete Protocol

Nano-Glo® Fluorofurimazine In Vivo Substrate Technical ManualPDF (1633 KB) – English

What's in the box?

Item	Part #	Size
Nano-Glo® Fluorofurimazine In Vivo Substrate	N410A	1 × 4.6μmol

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Patents and Disclaimers

Nano-Glo® Fluorofurimazine in vivo Substrate Limited Used Label License

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researchers may use this product for research use only, limited to animal imaging experiments or experiments performed to prepare for such animal imaging experiments; no commercial use is allowed. “Commercial use” means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; and (2) resale of the product, whether or not such product is resold for use in research. Researchers shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega. Notwithstanding the foregoing, researcher may use this product in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the product.

For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(b) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.

With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 11,691,976 and other patents pending.

What's in the box?

Item	Part #	Size
Nano-Glo® Fluorofurimazine In Vivo Substrate	N410A	5 × 4.6μmol

SDS

Search for SDS

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

Patents and Disclaimers

Nano-Glo® Fluorofurimazine in vivo Substrate Limited Used Label License

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researchers may use this product for research use only, limited to animal imaging experiments or experiments performed to prepare for such animal imaging experiments; no commercial use is allowed. “Commercial use” means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; and (2) resale of the product, whether or not such product is resold for use in research. Researchers shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega. Notwithstanding the foregoing, researcher may use this product in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the product.

For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(b) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.

With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.

U.S. Pat. No. 11,691,976 and other patents pending.

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