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Thedual glo luciferase assay system

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The Dual-Glo Luciferase Assay System is a laboratory equipment used to measure the activity of two different luciferase reporter enzymes simultaneously in a single sample. It provides a method for quantifying gene expression and evaluating cellular events.

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Lab products found in correlation

Multidrop combi dispenser, by Thermo Fisher Scientific (1 mentions) Viewlux plate reader, by PerkinElmer (1 mentions) Psv40 renilla, by Promega (1 mentions) Biotek synergy h4, by Agilent Technologies (1 mentions)

4 protocols using thedual glo luciferase assay system

1

High-throughput Luciferase Reporter Assay

Cited 1 time
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For each cell line tested, a total of 2000 cells per well in 4 µL
of media were dispensed using a Multidrop Combi dispenser (Thermo Fisher
Scientific) and a small cassette into barcoded 1536-well flat-bottom white
(Corning (Corning, NY, USA)) collagen-coated plates. After a 24 h
incubation, library compounds and controls were added to assay plates at a
volume of 23 nL/well via a NX-TR pintool station (Wako Scientific Solutions (San
Diego, CA, USA)), and the plates were incubated further (another 24 h).
Firefly and Renilla luminescence outputs were measured sequentially using the
Dual-Glo Luciferase Assay System (Promega) and the ViewLux plate reader
(PerkinElmer). The assay’s performance was stable throughout the screen.
The activity of each compound was normalized to control wells (DMSO alone),
which were included on each plate. Cells treated with DMSO alone were defined as
having 0% activity. Of note, the following libraries were screened: the
library of pharmacologically active compounds (LOPAC),13 (link) MIPE14 (link) and the
NIH Chemical Genomics Center (NCGC) Pharmaceutical Collection
(NPC).15
Bernstock J.D., Lee Y.J., Peruzzotti-Jametti L., Southall N., Johnson K.R., Maric D., Volpe G., Kouznetsova J., Zheng W., Pluchino S, & Hallenbeck J.M. (2015). A novel quantitative high-throughput screen identifies drugs that both activate SUMO conjugation via the inhibition of microRNAs 182 and 183 and facilitate neuroprotection in a model of oxygen and glucose deprivation. Journal of Cerebral Blood Flow & Metabolism, 36(2), 426-441.
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2

Luciferase Assay of miR-320c Inhibitor

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The HEK-293T cells which were seeded into the 48-well plates were treated with 5 pmol miR-320c inhibitor, inhibitor NC and 160 ng pSI-Check2-PTEN-3’UTR. After transfection for 48 h, the cells were collected and lysed. TheDual-Glo® Luciferase Assay System (Promega, USA) was utilized to determine the luciferase reporter activities. The experiment was repeated at least three times. The values of each well were determined as the ratio of Renilla luciferase to Firefly luciferase.
Sun Y., Qu H., Song Q., Shen Y., Wang L, & Niu X. (2022). High-glucose induced toxicity in HK-2 cells can be alleviated by inhibition of miRNA-320c. Renal Failure, 44(1), 1388-1398.
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3

Luciferase Assay for Odorant Receptor-I7 Activation

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The
Dual-Glo Luciferase Assay System (Promega) was used for the
luciferase assay as previously described.41 (link) Briefly, a plasmid encoding Rho-tagged rat OR-I7 (5 ng/well) was
transfected into the Hana3A cell line in 96-well plate format along
with plasmids encoding the human receptor trafficking protein, RTP1S40 (link) (5 ng/well), pSV40-Renilla (5
ng/well; Promega), CRE-luciferase (10 ng/well; Stratagene), and type
3 muscarinic acetylcholine receptor (M3-R)39 (2.5 ng/well). Then, 18 to 24 h following transfection, cells were
treated with compound 2 for 4 h at 37 °C, as described.39 Luminescence was measured using a Polarstar
Optima plate reader (BMG). Luciferase measurements were normalized
to Renilla luciferase measurements to control for
transfection efficiency and cell viability. Fold change values were
calculated by the formula (F1–F0)/F0, where F1 is the normalized luminescence response to
the odorant and F0 is the normalized luminescence
when no odorant was applied. Data were analyzed and the EC50 for 2 (≈10 μM) was estimated using Prism
5.0 and Microsoft Excel. Estimating the EC50s for the other
four odorants under the conditions of this assay was not possible
because they underwent significant evaporation. For this reason, we
used the GloSensor and calcium imaging assays described above to monitor
OR-I7 activation in real time.
Li Y., Peterlin Z., Ho J., Yarnitzky T., Liu M.T., Fichman M., Niv M.Y., Matsunami H., Firestein S, & Ryan K. (2014). Aldehyde Recognition and Discrimination by Mammalian Odorant Receptors via Functional Group-Specific Hydration Chemistry. ACS Chemical Biology, 9(11), 2563-2571.
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4

Cignal Transduction Pathway Profiling

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The Cignal Transduction 45-Pathway Reporter Array was purchased from SA Biosciences (Valencia, CA). Assays were performed according to the manufacturer's instructions. Luciferase activity was measured using The Dual-Glo® Luciferase Assay System (Promega, Madison, WI) with BioTek Synergy H4 multifunctional plate reader (BioTek).
Ranjan A., Iyer S.V., Ward C., Link T., Diaz F.J., Dhar A., Tawfik O.W., Weinman S.A., Azuma Y., Izumi T, & Iwakuma T. (2018). MTBP inhibits the Erk1/2-Elk-1 signaling in hepatocellular carcinoma. Oncotarget, 9(30), 21429-21443.
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