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One glo luciferase reagent

Manufactured by Promega
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One-Glo luciferase reagent is a luminescent assay reagent used for the detection and quantification of luciferase reporter gene activity in cell-based assays. The reagent contains the necessary components to lyse cells and produce a luminescent signal proportional to the amount of luciferase present in the sample.

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Lab products found in correlation

Viewlux plate reader, by PerkinElmer (4 mentions) Microplate spectrophotometer, by Molecular Devices (4 mentions) Topcount plate reader, by PerkinElmer (4 mentions) Vegf165a, by R&D Systems (3 mentions) Multidrop combi, by Thermo Fisher Scientific (2 mentions) Thermo scientific multidrop combi, by Thermo Fisher Scientific (2 mentions) Prism software version 8, by GraphPad (2 mentions) Reluc2p, by Promega (2 mentions) White 96 well plate, by Corning (2 mentions) Topflash, by Merck Group (2 mentions) Topflash, by Promega (2 mentions) Translt1 reagent, by Mirus Bio (2 mentions) Wallac victor2, by PerkinElmer (1 mentions) Oligofectamine, by Thermo Fisher Scientific (1 mentions) Synergy ht plate reader, by Agilent Technologies (1 mentions) Pherastar fs, by BMG Labtech (1 mentions) Celltiter fluor cell viability assay, by Promega (1 mentions) White 96 well plate, by SPL Life Sciences (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Nunc maxisorp 96 well plate, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

ONE-Glo reagent (61 citations) One-Glo (54 citations) Luciferase reagent (50 citations)

35 protocols using one glo luciferase reagent

1

HEK293 GLP-1R Luciferase Assay

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Example 23

HEK293 cells expressing surface GLP-1R and cAMP responsive luciferase reporter gene were seeded in 384 well plates at a density of 5000 cells per well. After 24 h incubation at 37° C. with 5% CO2, cells were treated with various concentrations of peptides and BLV1H12-GLP-1 clip fusion proteins and incubated for another 24 h. Subsequently, luciferase assay was performed using One-Glo luciferase reagent according manufacture's instruction (Promega). FIG. 20 and Table 16 show the activity of Ab-GLP-1 f fusion antibodies on HEK293 cells expressing GLP-1 receptor.

US10774132B2. Ultralong complementarity determining regions and uses thereof (2020-09-15). THE SCRIPPS RESEARCH INSTITUTE [US]. Inventors: Vaughn Smider [US], Omar A. Bazirgan [US], Hongyuan Helen Mao [US], Peter Schultz [US], Feng Wang [US], Yong Zhang [US].
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2

Evaluating GLP-1R Agonist Activity

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Example 15

HEK293 cells overexpressing surface GLP-1R and cAMP responsive luciferase reporter gene were seeded in 384 well plates at a density of 5,000 cells per well. After 24 h incubation at 37° C. with 5% CO2, cells were treated with various concentrations of exendin-4 peptide (SEQ ID NO: 228), trastuzumab (SEQ ID NOs: 19 and 22), trastuzumab-coil exendin-4 (SEQ ID NOs: 71 and 19), and trastuzumab-coil exendin-4 (SEQ ID NOs: 71 and 19) RN; and incubated for another 24 h. Subsequently, a luciferase assay was performed using One-Glo luciferase reagent according manufacture's instruction (Promega). FIG. 15 depicts a graphical representation of the data. The EC50 of exendin-4 was 0.03±0.004 nM. The EC50 of trastuzumab-coil exendin-4 was 3.8±0.2 nM. The EC50 of trastuzumab-coil exendin-4 RN was 0.01±0.001 nM.

US11673959B2. Coiled coil immunoglobulin fusion proteins and compositions thereof (2023-06-13). THE SCRIPPS RESEARCH INSTITUTE [US]. Inventors: Feng Wang [US], Yong Zhang [US], Yan Liu [US], Peter G. Schultz [US].
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3

Evaluating Glucagon-Like Peptide-1 Receptor Agonist Potency

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Example 15

HEK293 cells overexpressing surface GLP-1R and cAMP responsive luciferase reporter gene were seeded in 384 well plates at a density of 5,000 cells per well. After 24 h incubation at 37° C. with 5% CO2, cells were treated with various concentrations of exendin-4 peptide (SEQ ID NO: 228), trastuzumab (SEQ ID NOs: 19 and 22), trastuzumab-coil exendin-4 (SEQ ID NOs: 71 and 19), and trastuzumab-coil exendin-4 (SEQ ID NOs: 71 and 19) RN; and incubated for another 24 h. Subsequently, a luciferase assay was performed using One-Glo luciferase reagent according manufacture's instruction (Promega). FIG. 15 depicts a graphical representation of the data. The EC50 of exendin-4 was 0.03±0.004 nM. The EC50 of trastuzumab-coil exendin-4 was 3.8±0.2 nM. The EC50 of trastuzumab-coil exendin-4 RN was 0.01±0.001 nM.

US10683353B2. Coiled coil immunoglobulin fusion proteins and compositions thereof (2020-06-16). THE SCRIPPS RESEARCH INSTITUTE [US]. Inventors: Feng Wang [US], Yong Zhang [US], Yan Liu [US], Peter G. Schultz [US].
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4

Luciferase Assay for NRF2 Activity

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The enzymatic activity of luciferase in the lysates of HEK293 cell cultures was used as readout for NRF2 transcription factors activity. Under standard and RNAi screening conditions, cells were lysed 48 h post transfection by the addition of 20 μL of One-Glo luciferase reagent (Promega). An aliquot of 25 μL was transferred to white 384-well plates and luminescence generated by luciferase activity was determined 30 min later on a Wallac Victor2 (Perkin Elmer) luminescence reader.
Schumacher F.R., Schubert S., Hannus M., Sönnichsen B., Ittrich C., Kreideweiss S., Kurz T, & Rippmann J.F. (2016). RNAi Screen for NRF2 Inducers Identifies Targets That Rescue Primary Lung Epithelial Cells from Cigarette Smoke Induced Radical Stress. PLoS ONE, 11(11), e0166352.
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5

Luciferase Knockdown Assay Protocol

Cited 1 time
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Luciferase knockdown assays were performed as described in literature (14 (link)), with a few modifications. HeLa cells were counted and seeded at a density of 10 000 cells/well in a 96-well plate and were incubated for 24 h at 37°C with 5% CO2 prior to the experiment. Subsequently, cells were washed once with serum-free DMEM media and then 80 μl of serum-free DMEM media was added. siRNA and control duplexes were diluted up to 20 μl with serum-free media and transfection reagent (Oligofectamine, Invitrogen) and added to the appropriate well (for a total of 100 μl) at increasing concentrations (0.16, 0.8, 4, 20 and 100 nM). Cells were incubated overnight (for a total of 24 h post-transfection). Then 50 μl of ONE-Glo luciferase reagent (Promega, USA) was added to each well and luminescence was measured and normalized to protein levels using a Biotek Synergy HT plate reader.
Habibian M., Harikrishna S., Fakhoury J., Barton M., Ageely E.A., Cencic R., Fakih H.H., Katolik A., Takahashi M., Rossi J., Pelletier J., Gagnon K.T., Pradeepkumar P.I, & Damha M.J. (2020). Effect of 2′-5′/3′-5′ phosphodiester linkage heterogeneity on RNA interference. Nucleic Acids Research, 48(9), 4643-4657.
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6

STAT3 Activation Assay in HEK293

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HEK293-SIE reporter cells stably expressing either WT or mutant Y640F STAT3 were dispensed to prepared 384-well drug plates at 10 000 cells/well using a MultiDrop Combi peristaltic dispenser (Thermo Fisher, Carlsbad, CA, USA). Before plating, IL6 was added directly to the cell suspension (100 ng/ml) of WT STAT3 expressing cell lines. The cells were incubated with the drugs for 6 h after which the ONE-Glo luciferase reagent (Promega) was added according to the manufacturer’s protocol. The luciferase signal was measured with a PHERAstar FS plate reader (BMG Labtech, Ortenberg, Germany).
Kuusanmäki H., Dufva O., Parri E., van Adrichem A.J., Rajala H., Majumder M.M., Yadav B., Parsons A., Chan W.C., Wennerberg K., Mustjoki S, & Heckman C.A. (2017). Drug sensitivity profiling identifies potential therapies for lymphoproliferative disorders with overactive JAK/STAT3 signaling. Oncotarget, 8(57), 97516-97527.
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7

Multiplexed Luciferase and Cell Viability Assay

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ERR or PGC/ERR reporter HEK293 cells suspended in culture medium without puromycin were dispensed at 2500 cells/5 μL/well in tissue culture–treated 1536-well white assay plates (Greiner Bio-One North America, Monroe, NC, USA) using a Thermo Scientific Multidrop Combi (ThermoFisher Scientific, Inc.). Each compound has been tested at 15 concentrations ranging from 1.2 nM to 92 μM in the primary screening. After the cells were incubated at 37 °C with 5% CO2 for 6 h, 23 nL of compounds or control, XTC790, was transferred into the assay plates using a Wako Pintool station (Wako Automation, San Diego, CA, USA). The assay plates were incubated at 37 °C for 18 h, followed by the addition of 5 μL ONE-Glo luciferase reagent (Promega, Madison, WI, USA) using a Flying Reagent Dispenser (Aurora Discovery, Carlsbad, CA, USA). After 30 min of incubation at room temperature, the luminescence intensity of the assay plates was quantified using a ViewLux plate reader (PerkinElmer, Shelton, CT, USA). The assays were performed three times for each compound concentration. These ERR and PGC/ERR reporter assays were multiplexed with the CellTiter-Fluor Cell Viability Assay (Promega), a fluorescence-based cell viability assay, to assess compound cytotoxicity.
Lynch C., Zhao J., Sakamuru S., Zhang L., Huang R., Witt K.L., Merrick B.A., Teng C.T, & Xia M. (2019). Identification of Compounds That Inhibit Estrogen-Related Receptor Alpha Signaling Using High-Throughput Screening Assays. Molecules, 24(5), 841.
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8

SARS-CoV-2 Pseudovirus Neutralization Assay

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SARS-CoV-2 pseudotyped lentiviruses expressing full-length S proteins and HEK293T cells overexpressing human ACE2 (HEK293T/hACE2) were provided by the National RNAi Core Facility (Academia Sinica, Taiwan). The pseudovirus neutralization assays were performed using serial dilutions of RBD-mAbs pre-incubated with 1000 TU SARS-CoV-2 pseudovirus in 1% FBS DMEM for 1 h at 37 °C. The dilutions were incubated for 24 h at 37 °C in 96-well white plates (SPL Life Science), which had been pre-seeded with HEK293T/hACE2 cells at a density of 1 × 104 cells per well. The pseudovirus-containing culture medium was then replaced with 10% FBS DMEM for an additional 48 h. Next, ONE-Glo luciferase reagent (Promega) was added to each well for a 3 min incubation at 37 °C. Luminescence was measured using a microplate spectrophotometer (Molecular Devices) to determine pseudovirus infection efficacy. The half maximal inhibitory concentration (IC50) was calculated by nonlinear regression using Prism software version 8.1.0 (GraphPad Software Inc.).
Hsu F.F., Liang K.H., Kumari M., Chen W.Y., Lin H.T., Cheng C.M., Tao M.H, & Wu H.C. (2022). An efficient approach for SARS-CoV-2 monoclonal antibody production via modified mRNA-LNP immunization. International Journal of Pharmaceutics, 627, 122256.
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9

CD70 Activation Assay with Jurkat Cells

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Nunc MaxiSorp 96-well plates (Thermo Fisher # 44-2404-21) were coated overnight at 4oC with 1 μg/ml anti-human CD3 antibody (BD catalog # 555336) and dilutions of CD70 protein within 50 μl of PBS. The following day, plates were washed with PBS, and 50,000 Jurkat reporter cells were added in RPMI 1640 (Thermo Fisher # 72400047) with 10% FBS. The Jurkat reporter was stably transfected with an AP-1-dependent luciferase reporter. After a 4-h incubation at 37oC, One-Glo luciferase reagent (Promega # E6120) was added to the cells, incubated with gentle shaking for 5 min, and luminescence was measured by a Tecan Spark plate reader.
Liu W., Maben Z., Wang C., Lindquist K.C., Li M., Rayannavar V., Lopez Armenta I., Nager A., Pascua E., Dominik P.K., Oyen D., Wang H., Roach R.C., Allan C.M., Mosyak L, & Chaparro-Riggers J. (2021). Structural delineation and phase-dependent activation of the costimulatory CD27:CD70 complex. The Journal of Biological Chemistry, 297(4), 101102.
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10

Cytotoxicity and ADC Assays for CLDN18.2

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Bispecific cytotoxicity assays were performed in a 96-well plate format by mixing purified human CD3+ T cells and luciferase-labeled BxPC3/hCLDN18.2 or KATO-III/hCLDN18.2 cells, with an effector to target ratio (E:T) of 5:1. Serial dilutions of bispecific antibody were added to the plates and after 2 days of incubation, cell viability was assessed with the One-Glo luciferase reagent (Promega).
ADC assays were performed by plating BxPC3/hCLDN18.2 or KATO-III/hCLDN18.2 cells and adding serial dilutions of ADCs. After 4 days of incubation, viability of the cells was assessed by using CellTiter-Glo reagent (Promega).
Zhu G., Foletti D., Liu X., Ding S., Melton Witt J., Hasa-Moreno A., Rickert M., Holz C., Aschenbrenner L., Yang A.H., Kraynov E., Evering W., Obert L., Lee C., Sai T., Mistry T., Lindquist K.C., Van Blarcom T., Strop P., Chaparro-Riggers J, & Liu S.H. (2019). Targeting CLDN18.2 by CD3 Bispecific and ADC Modalities for the Treatments of Gastric and Pancreatic Cancer. Scientific Reports, 9, 8420.
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