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Dual lucifearase assay kit

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The Dual-Luciferase Assay kit is a laboratory reagent designed for the quantitative analysis of gene expression in biological samples. The kit provides a method for measuring the activities of two different luciferase reporter enzymes simultaneously within the same sample, allowing for normalized data analysis.

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Lab products found in correlation

Pgl4.10 luc2, by Promega (2 mentions) Fugene hd, by Promega (2 mentions) Q5 site directed mutagenesis kit, by New England Biolabs (2 mentions)

4 protocols using dual lucifearase assay kit

1

Luciferase Assay of Psmb11 Promoter

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The luciferase reporter assay was performed as described previously (2 (link)). In short, 4D6 cells were cotransfected with a renilla control plasmid (pRL Promega) and a luciferase reporter plasmid (pGL4.10[Luc2], Promega) under the control of a minimal wild-type Psmb11 promoter (designated β5t-luc) or a β5t promoter with a mutated FOXN1-binding sites (β5t-mut-luc) plus a FOXN1 construct of interest in a ratio of 1:10:10. Luciferease activity was measured 24 hours later using the Dual-Lucifearase Assay kit (Promega).
Rota I.A., Handel A.E., Maio S., Klein F., Dhalla F., Deadman M.E., Cheuk S., Newman J.A., Michaels Y.S., Zuklys S., Prevot N., Hublitz P., Charles P.D., Gkazi A.S., Adamopoulou E., Qasim W., Davies E.G., Hanson I., Pagnamenta A.T., Camps C., Dreau H.M., White A., James K., Fischer R., Gileadi O., Taylor J.C., Fulga T., Lagerholm B.C., Anderson G., Sezgin E, & Holländer G.A. (2021). FOXN1 forms higher-order nuclear condensates displaced by mutations causing immunodeficiency. Science Advances, 7(49), eabj9247.
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2

Foxn1 transcriptional regulation

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Promoter fragments of Psmb11 and Cd83 were subcloned into the pGL4.10(luc2, Promega) reporter plasmid and co-transfected with an expression vector encoding Foxn1 into HEK293 cells using FuGENE HD (Promega). Luciferease activity was measured 24hr later using the Dual-Lucifearase Assay kit (Promega). Motifs were mutated using Q5 Site-Directed Mutagenesis Kit (NEB).
Žuklys S., Handel A., Zhanybekova S., Govani F., Keller M., Maio S., Mayer C.E., Teh H.Y., Hafen K., Gallone G., Barthlott T., Ponting C.P, & Holländer G.A. (2016). Foxn1 regulates key target genes essential for T cell development in postnatal thymic epithelial cells. Nature immunology, 17(10), 1206-1215.
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3

Foxn1 transcriptional regulation

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Promoter fragments of Psmb11 and Cd83 were subcloned into the pGL4.10(luc2, Promega) reporter plasmid and co-transfected with an expression vector encoding Foxn1 into HEK293 cells using FuGENE HD (Promega). Luciferease activity was measured 24hr later using the Dual-Lucifearase Assay kit (Promega). Motifs were mutated using Q5 Site-Directed Mutagenesis Kit (NEB).
Žuklys S., Handel A., Zhanybekova S., Govani F., Keller M., Maio S., Mayer C.E., Teh H.Y., Hafen K., Gallone G., Barthlott T., Ponting C.P, & Holländer G.A. (2016). Foxn1 regulates key target genes essential for T cell development in postnatal thymic epithelial cells. Nature immunology, 17(10), 1206-1215.
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4

Luciferase Reporter Assay for Psmb11 Promoter

Check if the same lab product or an alternative is used in the 5 most similar protocols
The luciferase reporter assay was performed as described previously [2] . In short, 4D6 cells were co-transfected with a renilla control plasmid (pRL Promega), and a luciferase reporter plasmid (pGL4.10[Luc2], Promega) under the control of a minimal wild-type Psmb11 promoter (designated β5t-luc) or a β5t promoter with a mutated FOXN1 binding sites (β5t-mut-luc) plus a FOXN1 construct of interest in a ratio of 1:10:10. Luciferease activity was measured 24 h later using the Dual-Lucifearase Assay kit (Promega).
Rota I.A., Handel A.E., Klein F., Maio S., Dhalla F., Deadman M.E., Cheuk S., Newman J., Michaels Y.S., Žuklys S., Prevot N., Hublitz P., Charles P.D., Gkazi S.A., Adamopoulou E., Qasim W., Davies E.G., Hanson C., Pagnamenta A.T., Camps C., Dreau H., White A.J., James K.D., Fischer R., Gileadi O., Taylor J.C., Fulga T.A., Lagerholm B.C., Anderson G., Sezgin E., & Holländer G.A. (2021). A FOXN1 mutation competitively displaces wild-type FOXN1 from higher-order nuclear condensates to cause immunodeficiency.
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