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Nanoluc

Nanoluc is a novel bioluminescent reporter system derived from the deep-sea shrimp Oplophorus gracilirostris.
It offers enhanced luminescence, stability, and reduced size compared to traditional luciferase reporters.
Nanoluc has become a versatile tool for researchers across disciplines, enabling sensitive and quantitative analysis of gene expression, protein dynamics, and cellular processes.
PubCompare.ai leverages AI-driven analysis to help optimize Nanoluc protocols, identifying the most accurate and reproducible techniques from literature, preprints, and patents.
Experence the power of AI-driven protocol comparison to enhance your reasearch accuracy and productivity with Nanoluc.

Most cited protocols related to «Nanoluc»

1
Optimized Nluc Luciferase Assay
The buffer for Nluc reactions
consisted of 100 mM MES pH 6.0, 1 mM CDTA, 0.5% (v/v) Tergitol, 0.05%
(v/v) Mazu DF 204, 150 mM KCl, 1 mM DTT, and 35 mM thiourea. Furimazine
substrate was added to give a working reagent that was then added
in equal volume directly to assay samples (final concentration of
furimazine in the assay was commonly between 10 and 50 μM).
Complete methods and additional details can be found in the Supporting Information.
Hall M.P., Unch J., Binkowski B.F., Valley M.P., Butler B.L., Wood M.G., Otto P., Zimmerman K., Vidugiris G., Machleidt T., Robers M.B., Benink H.A., Eggers C.T., Slater M.R., Meisenheimer P.L., Klaubert D.H., Fan F., Encell L.P, & Wood K.V. (2012). Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate. ACS Chemical Biology, 7(11), 1848-1857.
Publication 2012
Biological Assay Buffers CDTA Tergitol Thiourea
2
Generation of SARS-CoV-2 Pseudotype Particles
To generate (HIV/NanoLuc) SARS-CoV-2 pseudotype particles, 5 × 106 293T cells were plated per 10-cm dish in 10 ml in growth medium. The following day, 7.5 µg pHIV-1NL4-3 ΔEnv-NanoLuc reporter virus plasmid and 2.5 µg SARS-CoV-2 or SARS-CoV plasmid (unless otherwise indicated, pSARS-CoV-2-SΔ19 was used) were mixed mix thoroughly with 500 µl serum-free DMEM (this represents a molar plasmid ratio of 1:0.55). Then, 44 µl polyethylenimine (PEI; 1 mg/ml) was diluted in 500 µl serum-free DMEM and mixed thoroughly.
To generate control virus lacking S, the S expression plasmid was omitted from the transfection, and the amount of PEI was reduced to 30 µl. The diluted DNA and PEI were then mixed thoroughly by pipetting or vortexing, incubated at 20 min at room temperature (RT), and added dropwise to the 293T cells. After 8-h or overnight incubation, the transfected cells were washed carefully twice with PBS and incubated in 10 ml DMEM++. At 48 h after transfection, the 10-ml supernatant was harvested, clarified by centrifugation at 300 g for 5 min, and passed through a 0.22-µm pore-size polyvinylidene fluoride syringe filter (Millipore; SLGVR33RS), aliquoted, and frozen at −80°C.
Schmidt F., Weisblum Y., Muecksch F., Hoffmann H.H., Michailidis E., Lorenzi J.C., Mendoza P., Rutkowska M., Bednarski E., Gaebler C., Agudelo M., Cho A., Wang Z., Gazumyan A., Cipolla M., Caskey M., Robbiani D.F., Nussenzweig M.C., Rice C.M., Hatziioannou T, & Bieniasz P.D. (2020). Measuring SARS-CoV-2 neutralizing antibody activity using pseudotyped and chimeric viruses. The Journal of Experimental Medicine, 217(11), e20201181.
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Publication 2020
Cells Centrifugation Culture Media Freezing HEK293 Cells Hyperostosis, Diffuse Idiopathic Skeletal Molar nanoluc Plasmids Polyethyleneimine polyvinylidene fluoride SARS-CoV-2 Serum Severe acute respiratory syndrome-related coronavirus Syringes Transfection Virus
3
SARS-CoV-2 Neutralization Assay
Five-fold serially diluted plasma from COVID-19 convalescent individuals and healthy donors or four-fold serially diluted monoclonal antibodies were incubated with the SARS-CoV-2 or SARS-CoV pseudotyped virus for 1 hour at 37°C degrees. The mixture was subsequently incubated with 293TACE2 cells for 48 hours after which cells were washed twice with PBS and lysed with Luciferase Cell Culture Lysis 5x reagent (Promega). Nanoluc Luciferase activity in lysates was measured using the Nano-Glo Luciferase Assay System (Promega) with Modulus II Microplate Reader User interface (TURNER BioSystems). Relative luminescence units obtained were normalized to those derived from cells infected with SARS-CoV-2 or SARS-CoV pseudotyped virus in the absence of plasma or monoclonal antibodies. The half-maximal inhibitory concentration for plasma (NT50) or monoclonal antibodies (IC50) was determined using 4-parameter nonlinear regression (GraphPad Prism).
Robbiani D.F., Gaebler C., Muecksch F., Lorenzi J.C., Wang Z., Cho A., Agudelo M., Barnes C.O., Gazumyan A., Finkin S., Hägglöf T., Oliveira T.Y., Viant C., Hurley A., Hoffmann H.H., Millard K.G., Kost R.G., Cipolla M., Gordon K., Bianchini F., Chen S.T., Ramos V., Patel R., Dizon J., Shimeliovich I., Mendoza P., Hartweger H., Nogueira L., Pack M., Horowitz J., Schmidt F., Weisblum Y., Michailidis E., Ashbrook A.W., Waltari E., Pak J.E., Huey-Tubman K.E., Koranda N., Hoffman P.R., West AP J.r., Rice C.M., Hatziioannou T., Bjorkman P.J., Bieniasz P.D., Caskey M, & Nussenzweig M.C. (2020). Convergent Antibody Responses to SARS-CoV-2 in Convalescent Individuals. Nature, 584(7821), 437-442.
Publication 2020
Biological Assay Cell Culture Techniques Cells COVID 19 Donors Luciferases Luminescence Monoclonal Antibodies nanoluc Plasma prisma Promega Psychological Inhibition SARS-CoV-2 Severe acute respiratory syndrome-related coronavirus Virus
4
SARS-CoV-2 Neutralization Assay
Five-fold serially diluted plasma from COVID-19 convalescent individuals and healthy donors or four-fold serially diluted monoclonal antibodies were incubated with the SARS-CoV-2 or SARS-CoV pseudotyped virus for 1 hour at 37°C degrees. The mixture was subsequently incubated with 293TACE2 cells for 48 hours after which cells were washed twice with PBS and lysed with Luciferase Cell Culture Lysis 5x reagent (Promega). Nanoluc Luciferase activity in lysates was measured using the Nano-Glo Luciferase Assay System (Promega) with Modulus II Microplate Reader User interface (TURNER BioSystems). Relative luminescence units obtained were normalized to those derived from cells infected with SARS-CoV-2 or SARS-CoV pseudotyped virus in the absence of plasma or monoclonal antibodies. The half-maximal inhibitory concentration for plasma (NT50) or monoclonal antibodies (IC50) was determined using 4-parameter nonlinear regression (GraphPad Prism).
Robbiani D.F., Gaebler C., Muecksch F., Lorenzi J.C., Wang Z., Cho A., Agudelo M., Barnes C.O., Gazumyan A., Finkin S., Hägglöf T., Oliveira T.Y., Viant C., Hurley A., Hoffmann H.H., Millard K.G., Kost R.G., Cipolla M., Gordon K., Bianchini F., Chen S.T., Ramos V., Patel R., Dizon J., Shimeliovich I., Mendoza P., Hartweger H., Nogueira L., Pack M., Horowitz J., Schmidt F., Weisblum Y., Michailidis E., Ashbrook A.W., Waltari E., Pak J.E., Huey-Tubman K.E., Koranda N., Hoffman P.R., West AP J.r., Rice C.M., Hatziioannou T., Bjorkman P.J., Bieniasz P.D., Caskey M, & Nussenzweig M.C. (2020). Convergent Antibody Responses to SARS-CoV-2 in Convalescent Individuals. Nature, 584(7821), 437-442.
Publication 2020
Biological Assay Cell Culture Techniques Cells COVID 19 Donors Luciferases Luminescence Monoclonal Antibodies nanoluc Plasma prisma Promega Psychological Inhibition SARS-CoV-2 Severe acute respiratory syndrome-related coronavirus Virus
5
Generation of SARS-CoV-2 Spike Protein Pseudotyped Virions
A plasmid expressing a C-terminally truncated SARS-CoV-2 S protein (pSARS-CoV2-Strunc) was generated by insertion of a human-codon optimized cDNA encoding SARS-CoV-2 S lacking the C-terminal 19 codons (Geneart) into pCR3.1. The S ORF was taken from “Wuhan seafood market pneumonia virus isolate Wuhan-Hu-1” (NC_045512). For expression of full-length SARS-CoV S protein, “Human SARS coronavirus Spike glycoprotein Gene ORF cDNA clone expression plasmid (Codon Optimized)” (here referred to as pSARS-CoV-S) was obtained from SinoBiological (Cat: VG40150-G-N). An env-inactivated HIV-1 reporter construct (pNL4-3ΔEnv-nanoluc) was generated from pNL4-331 (link) by introducing a 940 bp deletion 3’ to the vpu stop-codon, resulting in a frameshift in env. The human codon-optimized nanoluc Luciferase reporter gene (Nluc, Promega) was inserted in place of nucleotides 1–100 of the nef-gene. To generate pseudotyped viral stocks, 293T cells were transfected with pNL4-3ΔEnv-nanoluc and pSARS-CoV2-Strunc or pSARS-CoV-S using polyethyleneimine. Co-transfection of pNL4-3ΔEnv-nanoluc and S-expression plasmids leads to production of HIV-1-based virions carrying either the SARS-CoV-2 or SARS-CoV spike protein on the surface. Eight hours after transfection, cells were washed twice with PBS and fresh media was added. Supernatants containing virions were harvested 48 hours post transfection, filtered and stored at −80°C. Infectivity of virions was determined by titration on 293TACE2 cells. See also https://www.biorxiv.org/content/10.1101/2020.06.08.140871v1.
Robbiani D.F., Gaebler C., Muecksch F., Lorenzi J.C., Wang Z., Cho A., Agudelo M., Barnes C.O., Gazumyan A., Finkin S., Hägglöf T., Oliveira T.Y., Viant C., Hurley A., Hoffmann H.H., Millard K.G., Kost R.G., Cipolla M., Gordon K., Bianchini F., Chen S.T., Ramos V., Patel R., Dizon J., Shimeliovich I., Mendoza P., Hartweger H., Nogueira L., Pack M., Horowitz J., Schmidt F., Weisblum Y., Michailidis E., Ashbrook A.W., Waltari E., Pak J.E., Huey-Tubman K.E., Koranda N., Hoffman P.R., West AP J.r., Rice C.M., Hatziioannou T., Bjorkman P.J., Bieniasz P.D., Caskey M, & Nussenzweig M.C. (2020). Convergent Antibody Responses to SARS-CoV-2 in Convalescent Individuals. Nature, 584(7821), 437-442.
Publication 2020

Most recents protocols related to «Nanoluc»

1
NanoLuc Luminescence Activity Quantification
NanoLuc activity was quantified by measuring luminescence immediately following the addition of its substrate, fluorofurimazine, using a Synergy H1 plate reader. Protein samples for the assays were prepared by diluting free NanoLuc, Mx_pmut_NanoLuc, and Mx_wt_NanoLuc with Tris buffer (pH 7.5) to a final concentration of 50 pM with respect to NanoLuc. NanoLuc activity assays were initiated by adding serially diluted fluorofurimazine to the protein samples prepared in flat white bottom non-binding surface 96-well plates (Corning 3990). The final concentrations of fluorofurimazine ranged from 0.0977 μM to 12.5 μM, doubling in concentration at each dilution step. Luminescence readings were obtained in triplicate at room temperature, with a final reaction volume of 100 μL. As a background control, the same reaction mixtures without protein were used and subtracted from the luminescence signal of each sample. Non-linear regression curve analysis using the Michaelis–Menten model was conducted on substrate course initial rates using GraphPad Prism 9.
Kwon S., Andreas M.P, & Giessen T.W. (2024). Pore engineering as a general strategy to improve protein-based enzyme nanoreactor performance. bioRxiv.
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Publication Preprint 2024
2
Quantifying NanoLuc Luminescence in Cell Supernatants
NanoLuc concentrations in cell culture supernatants were quantified with the Nano-Glo Luciferase Assay System (N1110, Promega). 10 μL of cell culture supernatant was mixed with 10 μL of buffer/substrate mix (50:1) in 384-well plates (781076, Greiner Bio-One), which were briefly centrifuged at 1200 rpm and incubated for 5 min at room temperature. Luminescence intensity was measured using a Tecan Spark multiplate reader (Tecan AG).
Franko N., da Silva Santinha A.J., Xue S., Zhao H., Charpin-El Hamri G., Platt R.J., Teixeira A.P, & Fussenegger M. (2024). Integrated compact regulators of protein activity enable control of signaling pathways and genome-editing in vivo. Cell Discovery, 10, 9.
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Publication 2024
3
Purification of S-6P-NanoLuc Proteins
To express S-6P-NanoLuc proteins, Expi293 cells were transfected with S-6P-NanoLuc expression plasmids using ExpiFectamine 293 (Thermo Fisher Scientific). Four days later, the supernatant was harvested and loaded on Ni-NTA agarose, and after a thorough wash, S-6P-NanoLuc proteins were released after HRV 3C protease treatment.
Zhang F., Schmidt F., Muecksch F., Wang Z., Gazumyan A., Nussenzweig M.C., Gaebler C., Caskey M., Hatziioannou T, & Bieniasz P.D. (2024). SARS-CoV-2 spike glycosylation affects function and neutralization sensitivity. mBio, 15(2), e01672-23.
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Publication 2024
4
Quantification of nanoLuc mRNA and Protein
Naked nanoLuc mRNA capture and translation was assessed in murine primary cells (BMDCs) and two human epithelial cell lines (U-2 OS and MCF-7). To quantify nanoLuc mRNA levels and nanoLuc protein in BMDCs (Figure 5B and 5C), cells were seeded in 24 (250.000 cells/well) or 48 (120,000 cells/well) well plates, and cultured for 2 h to allow adhesion. Then, growth medium containing FBS was removed, cells were washed, incubated in growth medium without FBS and stimulated for 2 h with 1 μg / mL naked nanoLuc mRNA with 80 U / mL RI, thermally-inactivated RI, or without RI. To investigate the impact of varying concentrations of FBS and RI on nanoLuc translation by BMDCs (Figure D), cells were seeded in 48 well plates (120,000 cells/well) and cultured for 2 h to allow adhesion. Then, growth medium containing FBS was removed, cells were washed, incubated in growth medium containing varying concentration of FBS (0, 0.1, 1, 10%) and RI (0, 80, 120, 240, 320 U / mL) and stimulated with 1 μg / mL naked nanoLuc mRNA for 2 h. Subsequently, the medium containing the stimuli was removed and fresh growth medium with 10% FBS was added. After 6 h, nanoLuc mRNA was quantified by RT-qPCR. The RT-qPCR protocol was specifically designed to enable detection of full-length, undegraded nanoLuc mRNA. This was achieved by designing primers that selectively amplified the transcript 5’ end, which had been previously reverse-transcribed using oligo(dT) primers. After 24 h, nanoLuc protein translation in BMDCs was detected with Bio-Glo-NL Luciferase Assay System using a LUMIstar Optima luminometer. MCF-7 and U-2 OS cells were seeded in 48 well adherent plates. After reaching 80% confluence, growth medium was replaced with growth medium containing 0.1% FBS. Then, cells were incubated with varying concentration of nanoLuc mRNA (0, 0.25, 1, 4μg / mL), with either 80 U / mL RI or thermally inactivated RI for 2 h. To assess the involvement of dynamin-dependent endocytosis in naked nanoLuc mRNA capture, U-2 OS cells were incubated for 2 h with 80 μM Dynasore or DMSO, before addition of 1 μg / mL nanoLuc mRNA, with or without 80 U / mL RI. After 2 h, media was replaced with growth medium containing 10% FBS. Cells were cultured for additional 24 h, and nanoLuc protein levels were measured as previously described.
Castellano M., Blanco V., Calzi M.L., Costa B., Witwer K., Hill M., Cayota A., Segovia M, & Tosar J.P. (2024). Ribonuclease activity undermines immune sensing of naked extracellular RNA. bioRxiv.
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Publication Preprint 2024
5
Zoospore Cell Luminescence Assay
We collected zoospores released from electroporated cell populations, filtered them through a 25 mm Whatman Grade 1 filter paper and removed 10 µL to count the number of cells for normalization. We then pelleted the rest of the sample by centrifugation for 5 min at 2,500 × g at room temperature and resuspended the cell pellet in 50 µL of Bonner’s salts (10.27 mM NaCl, 10.06 mM KCl, 2.7 mM CaCl2 in ultrapure water). We then mixed the cell solution with 80 µL of NanoGlo luciferase assay reagent (Promega, Cat. No. N1110), transferred it to a 96-well plate well (PerkinElmer, Cat. No. 6005290), and measured luminescence using a FilterMax F5 microplate reader (Molecular Devices) every minute for 1 h using a 400 ms integration time. Reported luminescence values represent the mean of the measurements after 9, 10, and 11 min. Immediately after the plate reader measurement, we used a G:BOX iChemi XR gel imaging system (Syngene) to visualize NanoLuc luminescence in individual wells.
Kalinka E., Brody S.M., Swafford A.J., Medina E.M, & Fritz-Laylin L.K. (2024). Genetic transformation of the frog-killing chytrid fungus Batrachochytrium dendrobatidis. Proceedings of the National Academy of Sciences of the United States of America, 121(4), e2317928121.
Publication 2024

Top products related to «Nanoluc»

1
Nano glo luciferase assay system by Promega
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The Nano-Glo Luciferase Assay System is a luminescent reporter assay for the detection and quantification of luciferase activity in cell-based assays. The system utilizes a proprietary luciferase enzyme and optimized assay reagents to generate a robust, stable luminescent signal.
2
Nano glo dual luciferase reporter assay system by Promega
Sourced in United States, Japan
The Nano-Glo Dual-Luciferase Reporter Assay System is a laboratory product that measures the activity of two different reporter proteins simultaneously. It uses two distinct luciferase enzymes to generate bioluminescent signals, allowing for the analysis of multiple cellular events or pathways in a single experiment.
3
Nanoluc by Promega
Sourced in United States
NanoLuc is a bioluminescent reporter enzyme developed by Promega. It produces a bright, stable luminescent signal upon reaction with its substrate, enabling sensitive detection of biological events.
4
Passive lysis buffer (plb) by Promega
Sourced in United States, Germany, United Kingdom, France, Japan, China, Switzerland, Italy, Canada
Passive lysis buffer is a solution used for the gentle lysis of cells to extract proteins or other biomolecules. It facilitates the release of cellular contents without denaturing the target analytes.
5
Lipofectamine 2000 by Thermo Fisher Scientific
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Lipofectamine 2000 is a cationic lipid-based transfection reagent designed for efficient and reliable delivery of nucleic acids, such as plasmid DNA and small interfering RNA (siRNA), into a wide range of eukaryotic cell types. It facilitates the formation of complexes between the nucleic acid and the lipid components, which can then be introduced into cells to enable gene expression or gene silencing studies.
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Realtime glo mt cell viability assay by Promega
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The RealTime-Glo™ MT Cell Viability Assay is a luminescent-based assay that measures cell viability in real-time. It utilizes a Cell Viability Reagent that is added to the cell culture to produce a luminescent signal proportional to the number of viable cells.
7
Luciferase cell culture lysis 5 reagent by Promega
Luciferase Cell Culture Lysis 5× reagent is a concentrated solution that can be used to lyse cells and extract luciferase reporter proteins. It is designed for in vitro applications involving the measurement of luciferase activity in cell culture samples.
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Nano glo luciferase assay by Promega
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The Nano-Glo Luciferase Assay is a bioluminescent reporter assay system that utilizes a proprietary luciferase enzyme to quantify gene expression or protein interactions in a wide range of cell-based and biochemical applications.
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Fugene hd by Promega
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FuGENE HD is a non-liposomal transfection reagent designed for efficient delivery of DNA, RNA, and other macromolecules into a variety of cell types. It offers high transfection efficiency and low cytotoxicity.
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Nano glo substrate by Promega
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The Nano-Glo substrate is a luciferase-based reagent used for bioluminescent detection. It provides a robust luminescent signal that can be used to quantify target analytes in a variety of assay formats.

More about "Nanoluc"

Nanoluc is a cutting-edge bioluminescent reporter system derived from the deep-sea shrimp Oplophorus gracilirostris.
This innovative technology offers enhanced luminescence, stability, and a reduced size compared to traditional luciferase reporters, making it a versatile tool for researchers across disciplines.
Nanoluc enables sensitive and quantitative analysis of gene expression, protein dynamics, and cellular processes, revolutionizing the way scientists study these crucial biological phenomena.
PubCompare.ai, a powerful AI-driven platform, leverages advanced analysis to help researchers optimize Nanoluc protocols.
By scouring literature, preprints, and patents, PubCompare identifies the most accurate and reproducible Nanoluc techniques, empowering researchers to enhance their research accuracy and productivity.
Closely related to Nanoluc is the Nano-Glo Luciferase Assay System, a sensitive and versatile bioluminescent reporter system used to quantify gene expression, protein-protein interactions, and cell viability.
The Nano-Glo Dual-Luciferase Reporter Assay System, on the other hand, allows for the simultaneous measurement of two different reporter genes, providing valuable insights into cellular processes.
In addition to Nanoluc and Nano-Glo, researchers may utilize Passive lysis buffer, Lipofectamine 2000, RealTime-Glo™ MT Cell Viability Assay, Luciferase Cell Culture Lysis 5× reagent, Nano-Glo Luciferase Assay, FuGENE HD, and Nano-Glo substrate to further optimize their experimental workflows and enhance the accuracy and reproducibility of their findings.
Experience the power of AI-driven protocol comparison with PubCompare.ai and elevate your Nanoluc-based research to new heights of precision and productivity.