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Dual glow luciferase assay

Manufactured by Promega
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The Dual-Glow Luciferase Assay is a lab equipment product from Promega that measures the activity of two different luciferase reporter enzymes simultaneously in a single sample. It provides a sensitive and quantitative method for evaluating gene expression and reporter protein activity.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Xl site directed mutagenesis kit, by Qiagen (1 mentions) Lipo2000, by Thermo Fisher Scientific (1 mentions) Glomax 96 microplate luminometer, by Promega (1 mentions) Dharmafect duo, by Horizon Discovery (1 mentions) Psicheck 2 vector, by Promega (1 mentions) Quikchange 2 site directed mutagenesis kit, by Agilent Technologies (1 mentions) Phrl tk, by Promega (1 mentions) Synergy h1 hybrid multi mode microplate reader, by Agilent Technologies (1 mentions)

Close spelling variants of this product

Dual-Glo™ Stop and Glow Luciferase Assay system Dual-Glow Luciferase Assay kit Dual Glow Luciferase Assay System Promega dual glow assay Dual Luciferase Assay Luciferase assay

7 protocols using dual glow luciferase assay

1

Regulation of MCL1 by miR-125b

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MGC-803 cells (6 × 104) were seeded in 24-well plates immediately prior to transfection. The pMIR-MCL1 plasmids were transfected into MGC-803 cells using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. We also generated several inserts with deletions of 4 bp from the site of perfect complementarity of the MCL1 gene using the QIAGEN XL-site directed Mutagenesis Kit (QIAGEN, Valencia, CA). The miR-125b mimics and pMIR-MCL1 plasmids were cotransfected where indicated. Forty-eight hours after transfection, cells were assayed for both firefly and Renilla luciferase using the dual luciferase glow assay (Promega). Transfection experiments were performed in duplicate and repeated at least three times in independent experiments.
Wu S., Liu F., Xie L., Peng Y., Lv X., Zhu Y., Zhang Z, & He X. (2015). miR-125b Suppresses Proliferation and Invasion by Targeting MCL1 in Gastric Cancer. BioMed Research International, 2015, 365273.
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2

Mcl1 3'UTR Luciferase Assay

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MDA-MB-231 cells (6×104) were seeded in 24-well plates immediately prior to transfection. pMIR-Mcl1 and pMIR-Mcl1-mut were transfected into the MDA-MB-231 cells using Lipo2000 (Invitrogen) following the manufacturer's instructions. The miR-34a mimic was co-transfected where indicated. Cells were assayed for both firefly and renilla luciferase using the dual luciferase glow assay (Promega) 48 h post-transfection. Transfection experiments were performed in duplicate and repeated at least thrice in independent experiments.
Xiao X., Chen B., Liu X., Liu P., Zheng G., Ye F., Tang H, & Xie X. (2014). Diallyl Disulfide Suppresses SRC/Ras/ERK Signaling-Mediated Proliferation and Metastasis in Human Breast Cancer by Up-Regulating miR-34a. PLoS ONE, 9(11), e112720.
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3

Luciferase Assay for Mcl-1 Gene Regulation

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MDA-MB-435 cells (6×104) were seeded in 24-well plates immediately prior to transfection. The pMIR-Mcl1 and pMIR-Mcl1-mut constructs were transfected into MDA-MB-435 cells using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions. The miR-101 mimic was co-transfected where indicated. Forty-eight hours post-transfection, the cells were assayed for both firefly and renilla luciferase with the dual luciferase glow assay (Promega). Transfection experiments were performed in duplicate, and each experiment was repeated at least three times.
Liu X., Tang H., Chen J., Song C., Yang L., Liu P., Wang N., Xie X., Lin X, & Xie X. (2015). MicroRNA-101 inhibits cell progression and increases paclitaxel sensitivity by suppressing MCL-1 expression in human triple-negative breast cancer. Oncotarget, 6(24), 20070-20083.
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4

PD-L1-Promoter Driven Luciferase Assay

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Firefly luciferase expression controlled by PD-L1-promoter constructs was determined by triplicate measurements using the Dual-Glow luciferase assay (Promega, Mannheim, Germany). For normalization, constitutive Renilla-luciferase expression was determined thereafter.
Heckl S.M., Mau F., Senftleben A., Daunke T., Beckinger S., Abdullazade S., Schreiber S., Röcken C., Sebens S, & Schäfer H. (2021). Programmed Death-Ligand 1 (PD-L1) Expression Is Induced by Insulin in Pancreatic Ductal Adenocarcinoma Cells Pointing to Its Role in Immune Checkpoint Control. Medical Sciences, 9(3), 48.
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5

NF-κB Activation in NIH3T3 Cells

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NIH3T3 cells were plated in a 96 well plate, cultured to 70% confluence, and either transfected with 50 ng NF-κB plasmid or the corresponding control plasmid (all GeneCopoeia™/tebu-bio, Le Perray-en-Yvelines Cedex, France), both containing the firefly luciferase gene from Photinus pyralis. For determination of transfection efficacy, a plasmid containing the renilla gene from Renilla reniformis was used. 24 h after transfection, Dual-Glow-luciferase assay (GeneCopoeia™) was performed according to manufacturer’s instructions using a Glomax, 96 microplate luminometer from Promega Madison, WI, USA and obtained firefly activity was normalized to renilla activity. Data are shown as mean ± SD of at least 3 independent experiments.
Zhou Q., Einert M., Schmitt H., Wang Z., Pankratz F., Olivier C.B., Bode C., Liao J.K, & Moser M. (2016). MnTBAP increases BMPR-II expression in endothelial cells and attenuates vascular inflammation. Vascular pharmacology, 84, 67-73.
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6

Quantifying miR-142 Regulatory Targets

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Overexpression of miR-142 and luciferase sensor plasmids were performed as previously described (Fehniger et al., 2010 (link); Sullivan et al., 2012 (link)). The wild-type Socs1, Tgfbr1, and the first 1Kb of the Itgav 3’UTR were cloned into the psiCheck2 vector (Promega). The miR142-3p and −5p sites were mutated as indicated using the QuikChange II Site-Directed mutagenesis Kit (Agilent Technologies) following manufacturer’s instructions. miR-142-3p/5p pre-miRNA +/− 200bp flanking genomic sequence was sub-cloned into the pMND overexpression vector (a gift from M. Sands) (Fehniger et al., 2010 (link)). All primer sequences used for cloning are available upon request. 293T cells were co-transfected with 400ng of each vector using Dharmafect Duo (Dharmacon). After 48 hours, the Dual-Glow Luciferase Assay (Promega) was performed according to manufacturer’s instructions on an LD400 luminescence detector (Beckman Coulter). miR142 overexpression was confirmed by flow cytometry for GFP expression.
Berrien-Elliott M.M., Sun Y., Neal C., Ireland A., Trissal M.C., Sullivan R.P., Wagner J.A., Leong J.W., Wong P., Mah-Som A.Y., Wong T.N., Schappe T., Keppel C.R., Cortez V.S., Stamatiades E.G., Li M.O., Colonna M., Link D.C., French A.R., Cooper M.A., Boldin M.P., Reddy P, & Fehniger T.A. (2019). microRNA-142 is critical for the homeostasis and function of type-1 innate lymphoid cells. Immunity, 51(3), 479-490.e6.
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7

NF-κB Luciferase Assay for Ubiquitin Ligase Complex

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Method is described elsewhere (Ikeda et al., 2011; (link)Fennell et al., 2020) (link). Briefly, HEK293T were seeded in a 96-well plate and transfected with pNF-κB-Luc (Stratagene) and phRL-TK (Promega) as well as various HOIP, HOIL-1L and SHARPIN plasmids (as indicated) using GeneJuice. 48 hours post transfection, a Dual-glow luciferase assay (Promega E2940) was performed following the manufacturer's protocol. Synergy H1 hybrid multimode microplate reader (BioTek) was used to measure the luminescence signal. The luciferase signal from each sample was normalized to the corresponding Renilla luciferase signal in each well. All samples were normalized to the control samples transfected with an empty vector (pcDNA3.1-Myc), pNF-κB-Luc and phRK-TK.
Gómez-Díaz C., Jonsson G., Schodl K., Deszcz L., Bestehorn A., Eislmayr K., Almagro J., Kavirayani A., Fennell L.M., Hagelkrüys A., Kovarik P., Penninger J., & Ikeda F. (2021). The ubiquitin ligase HOIL-1L regulates immune responses by interacting with linear ubiquitin chains.
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