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Luciferase assay substrate kit

Manufactured by Promega
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The Luciferase Assay Substrate Kit is a laboratory product designed to measure luciferase enzyme activity. The kit provides the necessary reagents to quantify luciferase-based reporter gene expression in cell-based assays.

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Lab products found in correlation

Luminescent β galactosidase detection kit 2, by Takara Bio (2 mentions) Pcmv β gal plasmid, by Takara Bio (2 mentions) Pgl4.14 reporter plasmid, by Promega (2 mentions) Hygromycin, by Thermo Fisher Scientific (1 mentions) Cho tet on cells, by Takara Bio (1 mentions) Tet on system, by Takara Bio (1 mentions) Puromycin, by Merck Group (1 mentions) Celltiter glo luminescent cell viability assay, by Promega (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Luciferase assay substrate Luciferase Kit substrate Luciferase substrate Luciferase assay Promega luciferase kits Substrates

4 protocols using luciferase assay substrate kit

1

Regulation of IL17 Promoter Activity

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To examine the effect of PB on the activation of the Il17 promoter, Jurkat cells were co-transfected with pCMV-β-Gal plasmid (Clontech, Mountain View, CA), pCMV10-3xFlag-RORγ plasmid, and a pGL4.14 reporter plasmid (Promega Corp., Madison, WI, USA) under the control of human Il17-3kb-CNS promoter46 (link), and then treated with vehicle, or PB (0.5μmol/l), or GW9662 (1.0 μmol/l) for 1 h before PB incubation. After 24 h, cell extracts were lysed and the firefly luciferase and β-galactocidase activities were measured using a Luciferase Assay Substrate kit (Promega) and a Luminescent β-galactosidase Detection kit II (Clontech), respectively.
Yang Z., Liu M., Wang W., Wang Y., Cao B., Gao Y., Chen H, & Li T. (2017). Pseudolaric acid B attenuates atopic dermatitis-like skin lesions by inhibiting interleukin-17-induced inflammation. Scientific Reports, 7, 7918.
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2

Transient Transfection and TGF-β1 Signaling

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AGS cells were transiently transfected with SBE4-Luc, 3TP-Lux, ARE-Luc together
with forkhead activin signal transducer (FAST)-1, BRE-Luc and the internal
control pCMV-β-gal in 24-well plate using PEI reagent. After 24 h
transfection, cells were treated with 5 ng/ml TGF-β1 for 16 h.
In case of H. pylori infection experiment, cells were infected
with the indicated amount of H. pylori in 4 h before
TGF-β1 treatment. Luciferase activity was quantified by using Luciferase
Assay Substrate Kit (Promega Corp., Madison, WI). Values were normalized with
the β-galactosidase activity. All experiments were performed in triplicate
and repeated at least three times.
Nguyen T.T., Kim S.J., Park J.M., Hahm K.B, & Lee H.J. (2015). Repressed TGF-β signaling through CagA-Smad3 interaction as pathogenic mechanisms of Helicobacter pylori-associated gastritis. Journal of Clinical Biochemistry and Nutrition, 57(2), 113-120.
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3

Establishing Inducible ROR Cell Lines

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Doxycycline-inducible ROR stable cell lines were generated by transfecting a pTRE2 expression vector (Clontech, Mountain View, CA, USA) containing RORα or RORγ, into CHO Tet-on cells (Clontech) followed by transfection with the pGL4.27 luciferase (LUC) reporter vector (Promega, Madison, WI, USA) driven by 5xRORE. Single clones of pGL4.27-5xRORE- and pTRE2-ROR-expressing cells were selected from a medium containing hygromycin (Invitrogen, Grand Island, NY, USA) and puromycin (Sigma–Aldrich), respectively. CHO Tet-on cell lines were cultured in F12 medium supplemented with 10% FBS, and suitable for use in the Tet-on system (Clontech). To induce ROR expression, cells were treated for 20 h with 1 μM doxycycline in the presence or absence of a dilution series of the isoflavones. All the assays were performed in triplicate and repeated independently at least twice. RORE-mediated activation of the LUC reporter was measured with a Luciferase Assay Substrate Kit (Promega). cAMP-based cell viability was evaluated by CellTiter-Glo Luminescent Cell Viability Assay (Promega).
Kojima H., Takeda Y., Muromoto R., Takahashi M., Hirao T., Takeuchi S., Jetten A.M, & Matsuda T. (2015). Isoflavones enhance interleukin-17 gene expression via retinoic acid receptor-related orphan receptors α and γ. Toxicology, 329, 32-39.
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4

Isoflavone Regulation of IL-17a Promoter

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Jurkat cells were co-transfected with a pCMV-β-Gal plasmid (Clontech), pCMV10-3xFlag-RORα or pCMV10-3xFlag-RORγ plasmid, and a pGL4.14 reporter plasmid (Promega) under the control of human Il17a-3kb-CNS promoter (Zhang et al., 2012 (link)), using Lipofectamine 2000 (Invitrogen), and then treated with the vehicle, or 0.1, 1, 10 μM of the isoflavones. After 24 h, the firefly luciferase and β-galactocidase activities were measured using a Luciferase Assay Substrate Kit (Promega) and a Luminescent β-galactosidase Detection Kit II (Clontech), respectively. The firefly luciferase activity was normalized against β-galactocidase activity. All transfections were performed in triplicate and repeated at least twice.
Kojima H., Takeda Y., Muromoto R., Takahashi M., Hirao T., Takeuchi S., Jetten A.M, & Matsuda T. (2015). Isoflavones enhance interleukin-17 gene expression via retinoic acid receptor-related orphan receptors α and γ. Toxicology, 329, 32-39.
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