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One glo luciferase assay system

Manufactured by Promega
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Sourced in United States, Germany, Sweden

The ONE-Glo Luciferase Assay System is a luminescent reporter assay kit designed for quantitative measurement of firefly luciferase activity in cell-based assays. The assay is based on the luciferase enzyme, which catalyzes the oxidation of luciferin in the presence of ATP, Mg2+, and oxygen, resulting in the emission of light. The ONE-Glo Luciferase Assay System provides a sensitive and robust method for detecting and quantifying luciferase reporter gene expression.

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Lab products found in correlation

Lipofectamine 3000, by Thermo Fisher Scientific (17 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (15 mentions) Glo lysis buffer, by Promega (7 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions) Luminoskan ascent, by Thermo Fisher Scientific (6 mentions) Luminescence plate reader, by Berthold Technologies (5 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (4 mentions) Nano glo live cell assay system, by Promega (4 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Enspire multimode plate reader, by PerkinElmer (3 mentions) Lumat lb 9507, by Berthold Technologies (3 mentions) Jetprime dna and sirna transfection reagent, by Avantor (3 mentions) Fugene hd transfection reagent, by Promega (3 mentions) Glomax multi detection system, by Promega (3 mentions) Dual luciferase reporter assay system, by Promega (3 mentions) Envision 2104 multilabel reader, by PerkinElmer (3 mentions) Tristar2 lb942 luminometer, by Berthold Technologies (3 mentions) Bright glo luciferase assay system, by Promega (3 mentions) Opti mem, by Thermo Fisher Scientific (3 mentions) Celltiter glo luminescent cell viability assay, by Promega (3 mentions)

250 protocols using one glo luciferase assay system

1

Luciferase Assay for AP-1 Activation

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1 X 105 cells per each well were cultured in 48-well plate and incubated overnight. 1μg AP-1 luciferase reporter vector was transfected to cells by using 1ul lipofectamine, a transfection reagent, per well with OPTI-MEM and serum-free media (OPTI-MEM-SFM, 1:50 mixture). After 4 h, the media was changed to DEME supplemented with 5%. After 20h, cells were treated with dasatinib (200 nM) was pretreated for 30 min before 4-HNE stimulation. Cells were lysed by 100 μl ONE-Glo Luciferase Assay System (Promega, Corporate)’s lysis buffer per well. And lysate is transferred to 96-white well and 50 μl ONE-Glo Luciferase Assay System (Promega, Corporate)’s substrate solution was added. Level of Luciferase expression was measured by a luminometer (Berthold)
Jang E.J., Jeong H.O., Park D., Kim D.H., Choi Y.J., Chung K.W., Park M.H., Yu B.P, & Chung H.Y. (2015). Src Tyrosine Kinase Activation by 4-Hydroxynonenal Upregulates p38, ERK/AP-1 Signaling and COX-2 Expression in YPEN-1 Cells. PLoS ONE, 10(10), e0129244.
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2

STAT3-Luciferase Assay for Drug Sensitivity

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KR158 STAT3-luciferase (luc) reporter cells were seeded at a density 2 × 105 and grown in 6-well culture dishes. Cells were then left untreated or treated with 200 μM OXA, cisplatin (CDDP), bis-chloroethylnitrosourea (BCNU), or TMZ unless otherwise indicated for either 9 h or the length of time indicated. All drugs were dissolved in DMEM. Cells were pelleted by spinning in a microcentrifuge at 2000 rpm for 3 min in clear microcentrifuge tubes and lysed using the One-Glo Luciferase Assay System (Promega). Luciferase activity was determined using the One-Glo Luciferase Assay System (Promega) using a Glomax 20/20 luminometer.
Roberts N.B., Alqazzaz A., Hwang J.R., Qi X., Keegan A.D., Kim A.J., Winkles J.A, & Woodworth G.F. (2018). Oxaliplatin disrupts pathological features of glioma cells and associated macrophages independent of apoptosis induction. Journal of neuro-oncology, 140(3), 497-507.
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3

PPARα and FGF21 Luciferase Assay

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For the PPARα or FGF21 luciferase assay, 1.5 × 104 Ac2F cells/well were grown in a 96-well plate in DMEM supplemented with 10% FBS. The peroxisome proliferator response element (PPRE)-X3-TK-LUC plasmid (0.1 μg) (a kind gift from Dr. Christoper K. Glass, University of California, San Diego, CA, USA) and 0.01 μg of PPARα expression vector or pGL3B-Fgf21-LUC were transfected with Lipofectamine 3000 (0.1 μL) and P3000 (0.2 μL, Invitrogen) complexes in Opti-MEM (Invitrogen) according to the manufacturer’s instructions. The pcDNA empty vector was added to obtain an equal amount of plasmid DNA per transfection. After 24 h of transfection, cells were treated with vitamin C. Luciferase activity was tested using the ONE-Glo Luciferase Assay System (Promega, Madison, WI, USA) and a luminescence plate reader (Berthold Technologies GmbH & Co., Bad Wildbad, Germany).
Lee B., An H.J., Kim D.H., Lee M.K., Jeong H.H., Chung K.W., Go Y., Seo A.Y., Kim I.Y., Seong J.K., Yu B.P., Lee J., Im E., Lee I.K., Lee M.S., Yamada K.I, & Chung H.Y. (2022). SMP30-mediated synthesis of vitamin C activates the liver PPARα/FGF21 axis to regulate thermogenesis in mice. Experimental & Molecular Medicine, 54(11), 2036-2046.
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4

Lipid Nanoparticle Formulation and Characterization

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2-(aminomethyl)butanedioic acid hydrochloride was purchased from Enamine (Monmouth Jct., NJ). Dimethyl Formamide (DMF), Di-tert-butyl dicarbonate (BOC anhydride), Triethylamine (TEA), 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), N,N-Diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA) N,N’-Di-Boc-L-histidine dicyclohexylammonium salt, Nalpha,Nepsilon-Di-Boc-L-lysine and cholesterol were purchased from Fischer Scientific (Pittsburg, PA). 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol) (DSPE-mPEG(2000)) was purchased from Nanocs Inc (New York, NY). 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was purchased from Avanti Polar Lipids. Luciferase siRNA (5’-CUUACGCUGAGUACUUCGAtt-3’), human IKKα siRNA (5′-GGAGAUCUCCGAAAGCUGCtt-3′), human IKBKE siRNA (5′-GGUCUUCAACACUACCAGCtt-3′), mouse IKBKE siRNA (5’-GGUCUUCAACUCAGCCAGCtt-3) and Lipofectamine® 2000 were ordered from Invitrogen (Carlsbad, CA). 2-(p-toluidino)-6-naphthalene sulfonic acid (TNS) was obtained from Sigma Aldrich. CellTiter-Glo luminescent cell viability assay kit and ONE-Glo™ luciferase assay system were purchased from Promega (Madison, WI)
Palacios J.L., Zaror I., Martínez P., Uribe F., Opazo P., Socías T., Gidekel M, & Venegas A. (2001). Subset of Hybrid Eukaryotic Proteins Is Exported by the Type I Secretion System of Erwinia chrysanthemi. Journal of Bacteriology, 183(4), 1346-1358.
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5

Luciferase-based NF-κB activation assay

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Jurkat cells stably expressing human OX40 and NF-κB-luciferase construct and equal number of Raji cells were mixed and co-incubated with serial dilutions of BAT6026, PF-04518600 antibody or an control IgG1 for 5h at 37°C. Then the luciferase activity of samples were determined by a SpectraMax reader following the user guide of the ONE-Glo™ Luciferase Assay System (Promega).
Liang S., Zheng D., Liu X., Mei X., Zhou C., Xiao C., Qin C., Yue H., Lin J., Liu C., Li S, & Yu J.C. (2023). BAT6026, a novel anti-OX40 antibody with enhanced antibody dependent cellular cytotoxicity effect for cancer immunotherapy. Frontiers in Oncology, 13, 1211759.
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6

Pseudotyped HIV-1 infection in MM-6 cells

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For pseudotyped HIV-1 infection, 2.5 × 105 MM-6 cells were differentiated by 50 nM of PMA for 3 days in 12-well plate. In RNAi experiments, cells were transfected with 300 nM of indicated siRNA by Nucleofector Kit-V and suspended in PMA containing medium. After 3 days, NL4-3 viral solution (p24, 50 ng) was added, and incubated for 2 h. Following washing twice with pre-warmed medium, cells were cultured in PMA containing medium for additional 1 or 2 days, for neutral comet assay or Alu-gag two-step nested qPCR analysis, respectively. For luciferase assay, cell cultures (50,000 cells/well, in 96well plate) at 3 days post-infection (dpi) were directly lysed in equal volume of One-Glo Luciferase assay system (Promega) solution, and relative light units (RLU) were measured by microplate luminometer (Veritas). For FCM analysis of EGFP, cells were fixed with 1% paraformaldehyde at least 30 min at room temperature at 3dpi, and analysed by FACSCalibur on at least 10,000 cells.
Iijima K., Kobayashi J, & Ishizaka Y. (2018). Structural alteration of DNA induced by viral protein R of HIV-1 triggers the DNA damage response. Retrovirology, 15, 8.
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7

Measuring NFAT Translocation with Luciferase

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D1 nuclear factor of activated T cells translocation-firefly luciferase reporter cells were generated by transducing the D1 cells with Cignal Lenti NFAT Reporter with firefly luciferase (SABiosciences, Qiagen, Venlo, Limburg, Netherlands). NFAT nuclear translocation was detected by ONE-Glo™ Luciferase assay System (Promega, Madison, WI, USA) and the luminescence signal quantified with the GloMax®-Multi Detection System Luminometer module (Promega, Madison, WI, USA).
Wong A.Y., Oikonomou V., Paolicelli G., De Luca A., Pariano M., Fric J., Tay H.S., Ricciardi-Castagnoli P, & Zelante T. (2018). Leucine-Rich Repeat Kinase 2 Controls the Ca2+/Nuclear Factor of Activated T Cells/IL-2 Pathway during Aspergillus Non-Canonical Autophagy in Dendritic Cells. Frontiers in Immunology, 9, 210.
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8

Quantifying HIF Activity in AgNP-Treated MCF7 Cells

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MCF7 cells (grown in a 10 cm tissue culture dish) were transfected with 5 μg of HRE-dependent luciferase reporter construct pGL4.42 or a control construct pGL3-promoter (Promega Corporation, Madison, USA) using the Trans-Fast™ Reagent (Promega Corporation) according to the manufacturer’s instructions (pGL4.42 and pGL3 vector maps are provided in Figure S1). After 24 hours, the transfected cells were disassociated and plated in 96-well plates (5×103 cells/well). The cells were incubated for 3 hours to be allowed to adhere before being treated with the indicated concentrations of AgNPs (0, 20, 40, 60, 80, and 100 μg/mL in medium). HIF activity was induced by incubating the cells in hypoxic (0.1% O2) conditions or by treating the cells with cobalt chloride (CoCl2, 250 μM) for 16 hours. The luciferase activity was measured after 2 minutes by adding 100 μL of ONE-Glo™ luciferase assay system (Promega Corporation) according to the manufacturer’s instructions using a Multimode Microplate Reader (Berthold Technologies, Bad Wildbad, Germany).
Yang T., Yao Q., Cao F., Liu Q., Liu B, & Wang X.H. (2016). Silver nanoparticles inhibit the function of hypoxia-inducible factor-1 and target genes: insight into the cytotoxicity and antiangiogenesis. International Journal of Nanomedicine, 11, 6679-6692.
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9

Chimeric receptor signaling in Jurkat cells

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The mCD28/hCD28 and mCD28/hCD112R chimeras were generated by PCR and cloned into a pcDNA3.1(−) expression vector. We transduced chimera genes into Jurkat cells stably expressing a luciferase reporter under the control of NFAT response element (Jurkat-NFAT-Luc; Promega). Transfectants were selected with Zeocin and further enriched by flow cytometry sorting. Transfected Jurkat cells were stimulated with coated human CD3 mAb (OKT3) for 4 h with or without mouse CD28 mAb (clone 37.51). The presence of mouse CD28 mAb acts as an agonist to amplify signals transduced by the intracellular domain of the chimeras. After stimulation, cells were lysed with the ONE-Glo Luciferase Assay System (Promega) and measured for luminescent signal instantly.
Zhu Y., Paniccia A., Schulick A.C., Chen W., Koenig M.R., Byers J.T., Yao S., Bevers S, & Edil B.H. (2016). Identification of CD112R as a novel checkpoint for human T cells. The Journal of Experimental Medicine, 213(2), 167-176.
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10

Secosteroid-Induced Cytotoxicity Assay

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HaCaT, Caco-2, and Jurkat cells were selected for 1 week by culturing in medium containing additional 1.0 μg/mL puromycin. Each cell line was then plated in a 96-well plate (10000 cells/well, 100 μL medium) using FBS-free media and incubated for 24 h to synchronize the cells. Secosteroids at a series of concentrations were added separately to 96-well plate (1.0 μL/well). The final concentration of DMSO was 0.1%. Cells were incubated for another 24 h, and then 100 μL of solution of ONE-Glo luciferase assay system (Promega, Madison, WI) was added to each well. After 5 min reaction at room temperature, the luciferase signal was detected by a BioTek Synergy HT microplate reader (BioTek Instruments, Inc., Winooski, VT, US). All concentrations of secosteroids were tested in triplicate.
Lin Z., Marepally S.R., Ma D., Kim T.K., Oak A.S., Myers L.K., Tuckey R.C., Slominski A.T., Miller D.D, & Li W. (2016). Synthesis and Biological Evaluation of Vitamin D3 Metabolite 20S,23S-Dihydroxyvitamin D3 and Its 23R Epimer. Journal of medicinal chemistry, 59(10), 5102-5108.
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